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Planar Lipid Layers BLMs and their Application 1st
Edition H.T. Tien Digital Instant Download
Author(s): H.T. Tien, A. Ottova-Leitmannova
ISBN(s): 9780444509406, 0444509402
Edition: 1
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Year: 2003
Language: english
PLANAR LIPID BILAYERS (BLMs)
AND THEIR APPLICATIONS
Membrane Science and Technology Series

Volume 1 : Pervaporation Membrane Separation Processes

Edited by R.Y.M. Huang (1991)
Volume 2 : Membrane Separations Technology, Principles and Applications
Edited by R.D. Noble and S.A. Stern (1995)
Volume 3 : Inorganc Membranes for Separation and Reaction
By H.P. Hsieh (1996)
Volume 4 : Fundamentals of Inorganic Membrane Science and Technology
Edited by A.J. Burggraaf and L. Cot (1996)
Volume 5 : Membrane Biophysics
Edited by H. Ti Tien and A. Ottova-Leitmannova (2000)
Volume 6 : Recent Advances in Gas Separation by Microporous Ceramic Membranes
Edited by N.K. Kanellopoulos (2000)
Volume 7 : Planar Lipid Bilayers (BLMs) and their Applications
Edited by H.T. Tien and A. Ottova-Leitmannova (2003)
Membrane Science and Technology Series, 7

PLANAR LIPID BILAYERS (BLMs]

AND THEIR/~PL|CATIONS

Edited by

H.T. Tien
Department of Physiology, Michigan State University
East Lansing, M148824, USA

A. Ottova-Leitmannova
Department of Physiology, Michigan State University, East Lansing, M148824, USA
and
Center for Interface Sciences, Microelectronics Department, Faculty of Electrical
Engineering & Information Technology, Slovak Technical Universi~ 812 19 Bratislava,
Slovak Republic

2003

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 PREFACE

About four decades ago at a symposium on the Plasma Membrane,

organized jointly by the American and New York Heart Association, Donald
O. Rudin, Director of Basic Research at Eastern Pennsylvania Psychiatric
Institute, in Philadelphia, and his associates reported in a paper entitled
'Reconstitution of Cell Membrane Structure in vitro and its Transformation
into an Excitable System '. As evidenced by the present volume, the rest as
they say was history. The reconstituted system has been known under
various names (black, bimolecular, bilayer lipid membrane, BLM for short,
or simply planar lipid bilayer). Call whatever name you prefer, a
conventional BLM about 5 nm thick is interposed between two aqueous
solutions. It is, together with lipid vesicles (liposomes), the most widely
used experimental model of biomembranes. This liquid-crystalline BLM,
embodied in the lipid bilayer principle of biomembranes, and upon
numerous years of development, above and beyond performing as a physical
boundary, has been evolved, to serve:

9 as a discriminatory barrier
9 as a conduit for transport
9 as a reactor for energy conversion
9 as a transducer for signal processing
9 as a bipolar electrode for redox reactions
9 as a site for molecular recognition and/or
9 other diverse functions such as apoptosis, signal transduction, etc.

To mark BLM's 4 0 th anniversary, we have invited some of our

colleagues to contribute a chapter describing their unique approach. It was
impossible to invite all of the active researchers of planar lipid bilayers, past
and present, to put in their work to this volume. The Editors offer their
apologies and will endeavor to include other investigators' pioneering and
outstanding work on planar BLMs and liposomes, not represented here, in
future volumes.

The Editors
This Page Intentionally Left Blank
 P L A N A R LIPID BILAYERS (BLMs) AND
THEIR APPLICATIONS
H.T. Tien and A. Ottova (Editors)

Preface

I. Lipid bilayer principle of b i o m e m b r a n e s

Chapter 1 The lipid bilayer concept: Experimental realization

and current applications
H.T. Tien and A. Ottova

Chapter 2 Dielectric and Electrical Properties of Lipid Bilayers in

Relation to their Structure
H.G.L. Coster 75

Chapter 3 Boundary poterrtials of bilayer lipid membranes: methods

and interpretations
Yu. A. Ermakov and V.S. Sokolov 109

Chapter 4 Effect of anisotropic properties of membrane constituents

on stable shapes of membrane bilayer structure
A. lglic and V. Kralj-lglic 143

Chapter 5 Elastic properties of BLMs and pore formation

D. Popescu, S. Ion, A. Popescu, and Liviu Movileanu 173

Chapter 6 Mechanoelectric properties of BLMs

A.G. Petrov 205

Chapter 7 Chain-ordering phase transition in bilayer: kinetic

mechanism and its physiochemical and physiological
implications
D. Kharakoz 239

Chapter 8 Coupling of chain melting and bilayer structure: domains,

rafts, elasticity and fusion
Th. Heimburg 269
 ooo

Pill Contents

Chapter 9 Water transport

P. Pohl 295

Chapter 10 Membrane-macromolecule interactions and their structural

consequences
S. May and A. Ben-Shaul 315

II. Ion Selectivity, Specificity, and M e m b r a n e R e c o n s t i t u t i o n

Chapter 11 Investigation of substrate-specific porin channels in BLMs

R. Benz 347

Chapter 12 Planar lipid bilayer analyses of bacterial porins"

the role of structure in defining function
M.A. Arbing and J.W. Coulton 371

Chapter 13 Reconstitution in planar lipid bilayers of ion

channels synthesized in ovo and in vitro
L.K. Lyford and R.L. Rosenberg 391

Chapter 14 Multi-channel and single-channel investigation of

protein and peptide incorporation into BLMs
E. Gallucci, S. Miceili, and V. Picciarelli 413

Chapter 15 Structure and function of plant membrane ion channels

reconstituted in planar lipid bilayers
M. Smeyers, M. L~onetti, E. Goormaghtigh, and
F. Hombl6 449

Chapter 16 Reconstituting SNARE proteins into BLMs

K.T. Rognlien and D.J. Woodbury 479

Chapter 17 Mitochondrial ion channels, their isolation and study in

planar BLMs
G. Mironova 489

Chapter 18 The use of liposomes to detect channel formation

mediated by secreted bacterial proteins
V. Cabiaux, S. Vande Weyer and J.M. Ruysschaert 517
Contents ix

Chapter 19 The quest for ion channel memory using a planar BLM
R. Cassia-Moura 539

Chapter 20 Symmetric and asymmetric planar lipid bilayers of various

lipid composition: a tool for studying mechanisms and
lipid specificity of peptide/membrane interactions
A. Wiese, T. Gutsmann, and U. Seydel 569

Chapter 21 Insights into ion channels from peptides in planar

lipid bilayers
H. Duelohier 589

Chapter 22 Permeation property and intramembrane environments

of synthetic phytanyl-chained glycolipid membranes
T. Baba, H. Minamikawa, M. Hato and T. l-Ianda 605

Chapter 23 Modulation of planar bilayer permeability by electric

fields and exogenous peptides
A. Gliozzi, F. Gambale and G. Menestrina 633

Chapter 24 Gravitational impact on ion channels incorporated

into planar lipid bilayers
M. Wiedemann, H. Rahmann and W. Hanke 669

Chapter 25 Advantages and disadvantages of patch-clamping versus

using BLM
M.L. Kelly and D.J. Woodbury 699

Chapter 26 Using bilayer lipid membranes to investigate the

pharmacology of intracellular calcium channels
P. Koulen 723

III. Planar B L M s in Biotechnology

Chapter 27 Systems aspects of supported membrane biosensors

I.R. Peterson and J.A. Beddow 735

Chapter 28 Biosensors from interactions of DNA with lipid membranes

U.J. KruU, D.P. Nikolelis and J. Zeng 767
x Contents

Chapter 29 Structure and electrochemistry of fullerene lipid-hybrid

and composite materials
Naotoshi Nakashima 789

Chapter 30 Analytical applications ofbilayer lipid membrane systems

Marek Trojanowicz 807

Chapter 31 Transmembrane voltage sensor

J.A. Cohen, B. Gabriel, J. Teissi6, M. Winterhalter 847

Chapter 32 Domains, cushioning and patteming ofbilayers by surface

interactions with solid substrates and their sensing properties
R. Vidu, T.V. Ratto, M.L. Longo, and Pieter Stroeve 887

Chapter 33 Supported planar lipid bilayers (s-BLMs, Sb-BLMs, etc.)

A. Ottova, V. Tvarozek, and H.T. Tien 917

IV. L i g h t - i n d u c e d p h e n o m e n a and spectroscopy

Chapter 34 Photoinduced charge separation in lipid bilayers

D. Mauzerall and K. Sun 963

Chapter 35 Photosynthetic pigmem-protein complexes in planar lipid

membranes
W.I. Gruszecki and A. Wardak 981

Chapter 36 Biochemical applications of solid supported membranes

on gold surfaces: quartz crystal microbalance and
impedance analysis
A. Janshoff, H.-J. Galla and C. Steinem 991

Chapter 37 Simultaneous measuremem of spectroscopic and

physiological signals from a planar bilayer system
Y. Hanyu 1017

Subject Index 1031

Planar Lipid Bilayers (BLMs) and their Applications
H.T. Tien and A. Ottova-Leitmannova (Editors)
9 2003 Elsevier Science B.V. All rights reserved.

Chapter 1

The lipid bilayer concept:

Experimental realization and current applications

H. T. Tien and A. Ottova

Physiology Department, Biomedical and Physical Sciences Building

Michigan State University, East Lansing, MI 48824 (USA)

1. INTRODUCTION

It may sound far fetched but it is nonetheless true that the concept of the lipid
bilayer of cell or biological membranes began with the observations of R. Hooke of
Hooke's law fame, who in 1672 coined the word 'cell' to describe the array of a
cork slice under a microscope he constructed. Using the microscope, Hooke
discovered 'black holes' in soap bubbles and films. Years later, Isaac Newton
estimated the blackest soap film to be about 3/8 x 10 -6 inch thick. Modem
measurements give thickness between 5 and 9 nm, depending on the soap solution
used.
Question" How does one embark from black soap films to black lipid membranes
(BLMs) as models of biological membranes? Here we must go back in time
to the early 1960s. A good starting point, perhaps, is the conference held in
New York City.

1.1. Symposium on the Plasma Membrane

In December 1961, at the meeting sponsored by the American and New York
Heart Association, when a group of unknown researchers reported the
reconstitution of a bimolecular lipid membrane in vitro, the account was met with
skepticism. Those present included some of the foremost proponents of the lipid
bilayer concept, such as Danielli, Davson, Stoeckenius, Adrian, Mauro, Finean, and
many others [1]. The research group led by Donald Rudin began the report with a
description of mundane soap bubbles, followed by 'black holes' in soap films, etc.
ending with an invisible 'black' lipid membrane, made from extracts of cow's
brains. The reconstituted structure (60-90 A thick) was created just like a cell
membrane separating two aqueous solutions. The speaker then said:
2 Chapter 1

"... upon adding one, as yet unidentified, heat-stable compound., from

fermented egg white ....... to one side o f the bathing solutions .... lowers the
resistance.., by 5 orders o f magnitude to a new steady state .... which changes with
applied potential... Recovery is prompt.., the phenomenon is indistinguishable...
from the excitable alga Valonia... , and similar to the frog nerve action potential... "

As one member of the amused audience remarked, "... the report sounded
like.., cooking in the kitchen, rather than a scientific experiment!" That was in
1961, and the first report was published a year later [2]. In reaction to that report,
Bangham, the originator of liposomes [3], wrote in a 1995 article entitled
'Surrogate cells or Trojan horses"

"... a preprint o f a paper was lent to me by Richard Keynes, then Head o f the
Department o f Physiology (Cambridge University), and my boss. This paper was a
bombshell... They (Mueller, Rudin, Tien and Wescott) described methods for
preparing a membrane ..... not too dissimilar to that o f a node o f Ranvier... The
physiologists went mad over the model, referred to as a 'BLM', an acronym for
Bilayer or by some for Black Lipid Membrane. They were as irresistible to play
with as soap bubbles."

Indeed, the Rudin group, then working on the 9 th floor of the Eastern
Pennsylvania Psychiatric Institute (now defunct) in Philadelphia, Pa., was playing
with soap bubbles with the 'equipment' purchased from the local toyshop [4].
While nothing unusual for the researchers at work, it must have been a curious and
mysterious sight for the occasional visitors who happened to pass through the
corridor of laboratories there! However, playing with soap bubbles scientifically
has a long, respectable antiquity, as already mentioned in the introductory
paragraph.

1.2. Origins of the Lipid Bilayer Concept

Nowadays it is taken for granted that the lipid bilayer comprises the
fundamental structure of all biomembranes. The recognition of the lipid bilayer as a
model for biomembranes dates back to the work of Gorter and Grendel published in
1925 [5] However, the origin of the lipid bilayer concept is much older, and is
traceable to black soap bubbles more than three centuries ago!
The early observation of 'black holes' in soap films by Hooke and Newton
had a profound influence on the development of the lipid bilayer concept of
biomembranes and its subsequent experimental realization in planar lipid bilayers
and spherical liposomes. In this connection, there has been much discussion lately
The lipid bilayer concept 3

on self-assemblies of molecules, meaning the aggregation of molecular moieties

into thermodynamically stable and more ordered structures. Without question, the
inspiration for these exciting developments comes from t h e biological world,
where, for example, Nature uses self-assembly, as a strategy to create complex,
functional structures, such as viral protein coatings, and DNA, besides the above-
mentioned lipid bilayer of cell membranes. Many researchers have reported self-
assembling systems such as Langmuir-Blodgett monolayers and multi-layers [6]. A
broad list of man-made, self-assembling systems involving amphiphilic molecules
is given in Table 1.

Table 1
Experimental self-assembling interfacial amphiphilic systems

System Interfaces
1. Soap films air]soap solution]air
2. Monolayers air]molecular layer]water
3. Micelles (water-in-oil) water ] monolayer ] oil
Micelles (oil-in-water) oil ] monolayer ] water
4. Multilayers air]molecular layers]water
5. Bilayers (BLMs and Liposomes) water ] BLM ] water
6. Nuclepore supported BLMs water [ BLM ] water
7. Gold supported Monolayers air]molecular layers]gold
8. Metal supported BLMs water]lipid bilayer ] metal
(s-BLMs)
9. Salt-bridge supported BLMs water]lipid bilayer [ hydrogel
(sb-BLMs)
10. Tethered BLMs (t-BLMs) gold [ SH-BLM ] water

Where the vertical line[ denotes an interface.

1.3. Early experimental evidence for the lipid bilayer

Prior to the seminal work of Gorter and Grendel on the red blood cells
(RBC), several pertinent questions had been raised:
9 Was there a plasma membrane (Pfeffer, 1877)?
9 What was the nature of the plasma membrane (Overton, 1890)?
9 What is the meaning o f membrane potential (Bernstein, 1900)?
9 How thick is the plasma membrane (Fricke, 1925)?
9 What is the molecular organization oflipids in the plasma membrane (Gorter
and Grendel, 1925)?
4 Chapter 1

Before answering the last posed question, let us digress for the moment to a
different topic, namely, interfacial and colloid chemistry. It is no exaggeration that
modern investigation of interfacial and colloid chemistry began in the kitchen sink!
Over a span of 4 years (1891-1894), Agnes Pockels (a house wife working over the
kitchen sink) reported in Nature (see Ref. [7]) how surface films could be enclosed
by means of physical barriers to about 20 A2/molecule. To be more accurate
historically, the first landmark is generally bestowed to Benjamin Franklin, who in
1774 demonstrated that a teaspoonful of oil (olive ?) is able to calm a half-acre (~
2000 m 2 ) surface of a pond. Pockels' observation was followed about a quarter of
century later by the quantitative investigation of Langmuir, using a setup now
known as Langmuir's film balance. Langmuir deduced the dimensions and
structures of fatty acid molecules at the air-water interface. Our current
understanding of the structure and function of biomembranes can be traced to the
studies of experimental interfacial systems such as soap bubbles and Langmuir
monolayers, which have evolved as a direct consequence of applications of
classical principles of colloid and interfacial chemistry [6]. Accordingly, interfaces
play a vial role in molecular biology, since most ligand-receptor interactions
occurring at surfaces and interfaces. In the 19th century, the first observations were
made that surfaces control biological reactions. The challenge is to fully develop
the biological model for surface/interface science in the highly complex and
interactive in vivo biological setting. Concerning these, surface and colloid
scientists have known for decades that amphiphilic compounds such as
phospholipids can self-organize or self-assemble themselves into supramolecular
structures of emulsions, micelles, and bilayer lipid membranes (planar BLMs and
spherical liposomes). Thermodynamics favors self-assembly, if the molecules of an
amphipathic phospholipid (e.g. phosphatidylcholine, PC) are in water, the
hydrocarbon chains will want to be 'away' from the environment. They could all go
to the top (like oil on water), or they could have the hydrocarbon chains point
toward each other. With the hydrocarbon chains pointing toward each other,
amphiphilic molecules of phospholipids could adapt at least three different
configurations (see Fig. 1). The three ways in which they do so are to form small
spherical micelles, liposomes or to form a lamellar BLM. A micelle is a small
structure, generally less than two gM in diameter, while a free-standing BLM
typically has a thickness of about 5 nm and can have an area of several square
millimeters or more. Mechanical treatment or sonication of lipids can produce
spherical BLMs (i.e. liposomes or lipid vesicles), which contain aqueous solution
enclosed within a lipid bilayer. Liposomes are considerably larger than micelles,
ranging from tenth of gM to cm in diameter [3].
The lipid bilayer concept 5

Studies of such BLMs have given us much insight into the functions and
operation of biomembranes in living cells. Unmodified BLMs are highly permeable
to water molecules, while ions such as Na + and K + can traverse them hardly at all
(by nine orders of magnitude lower!). As shown in Fig. 1, a micelle may be
depicted as a ball with the lipid polar groups on the outside and the hydrocarbon
chains pointing together, as in w/o (water-in-oil) micelles, and vice versa as in o/w
(oil-in-water) micelles. Interestingly, micelles are generally not formed by
phospholipids in aqueous solutions, since the fatty acyl hydrocarbon chains in
phosphoglycerides (PG), sphingomyelins (SM), and all glycolipids (GL), are too
large to fit into the interior of a micelle. Thus, naturally occurring phospholipids
have a rather low cmc (critical micelle concentration); it is on the order of 5 x 10.4
~tM. The arrangement of atoms and molecules at the surface or interface of a
substance differs from that in the bulk, owing to their surroundings.
Gibbs, Laplace and Young formulated the basic principles of interfacial
chemistry. Specifically Gibbs articulated the thermodynamics of interfaces,
whereas Young and Laplace introduced the concept of surface tension. It is thus
useful in many fundamental and practical applications such as molecular films,
biomembranes, and lipid-based biosensors to have some knowledge concerning the
behavior of substances at the interface. The nature of interfacial phenomena is
inherently interdisciplinary involving physics, chemistry, biology, and engineering
[6].
Now, answering the question posed earlier: "What is the molecular organization
o f lipids in the plasma membrane?" Shortly after Langmuir' s 1917 paper showed
that his simple apparatus could provide the data to estimate the dimensions of a
molecule, Gorter, a pediatrician and Grendel, a chemist, determined the area
occupied by lipids extracted from red blood cell (RBC) ghosts. They found that
there were enough lipids to form a layer two molecules thick over the whole cell
surface, that is

Surface area occupied (from monolayer experiment)

~2 (1)
Surface area o f RBC (from human, pig or rat source)

On this basis, Gorter and Grendel suggested that the plasma membrane of RBCs
might be thought of as a bimolecular lipid leaflet [5]; they wrote

"... o f which the polar groups are directed to the inside and to the outside, in
much the same way as Bragg supposes the molecules to be oriented in a 'crystal' o f
fatty acid, and as the molecules of a soap bubble are according to Perrin."
6 The lipid bilayer concept

Hence, the concept of a lipid bilayer as the fundamental structure of cell

membranes was born, and has ever since dominated our thinking about the
molecular organization of all biomembranes!
From the 1930s onward, research on biomembranes diverged into two major
directions: (i) the further elaboration of the lipid bilayer concept using increasingly
sophisticated physical chemical techniques including x-ray diffraction and electron
microscopy, and (ii) the invention of model membrane systems so that physical,
biochemical, and physiological processes may be isolated and analyzed in
molecular terms. The former approach resulted in many insightful cartoons,
prominently among these are the classic model of Harvey, Danielli and Davson, the
spherical micelle model of Sjostrand and of Lucy, the various other models
proposed by Benson, Lenard, Singer and Nicolson, and by Green, as well as the
Chop-Suey (Smorgasbord) model [6,7]. Some of these models are in essence an
elaboration of the 'unit membrane' hypothesis of Robertson, which led eventually
to the so-called fluid mosaic model of biomembranes. The other direction of
research culminated in the reconstitution of lipid bilayers in vitro [2,3], as described
in the following section.

0 EXPERIMENTAL REALIZATION OF THE BILAYER LIPID

MEMBRANES

How did the pioneering researchers get the idea in 1960 to form the first
experimental bilayer lipid membrane? In short, the early investigators conceived
the idea by playing with soap bubbles and films whose molecular organization
provided the key! What follows-is a personal account that might be of some historic
interest [ 1-7].
In the late 1950s, while a research group headed by Donald O. Rudin, was
investigating the ion specificity of lipid monolayers and multilayers (the Langmuir-
Blodgett type), two refreshing publications appeared, which exerted great influence
on the thinking of Rudin and his associates, and altered the course of their research.
The first was a reprint of C.V. Boys' classic book on soap bubbles. The second was
a volume dedicated to N.K. Adam, in which A.S.C. Lawrence recounted succinctly
the highlights in the development of monolayers, soap films, and colloid chemistry.
Lawrence's account in particular brought into focus the relationship among the
various topics. Most significant was the mention of Newton's observation of the
black soap films (described in detail in Boys' book). The confirmation of the
thickness of black soap films measured by several investigators, including Perrin
and Dewar, since Hooke and Newton's time, and the bimolecular leaflet model for
the plasma membrane suggested independently by Gorter and Grendel, and by
Chapter 1 , 7

Fricke in the 1920's, which was then being corroborated by numerous

investigations. The structure of black soap films led to the realization by Rudin and
his co-workers in 1960 that a soap film in its final stages of thinning has a structure
comprised of two fatty acid monolayers sandwiching an aqueous solution. With the
above background in mind, Rudin and his associates simply proceeded to make a
'black lipid bubble' under water; their effort was successful [2-4]. As far as
forming a 'black', bimolecular or bilayer lipid membrane (BLM) is concerned,
there is not a lot to it. It is worth stating, however, that this experimental realization
drew on three centuries of annotations! The preceding account, in essence, is the
historical origin of the bilayer lipid membrane (dubbed BLM).*
Rudin and his colleagues showed that, an under water 'lipid film', or a BLM
formed from brain extracts was self-sealing to puncture with many physical and
chemical properties similar to those of biomembranes. Upon modification with a
certain compound (called EIM, Excitability-Inducing-Molecule), this otherwise
electrically 'inert' structure became excitable, displaying characteristic features
similar to those of action potentials of the nerve membrane. An unmodified lipid
bilayer separating two similar aqueous solutions, on the order of 5 nm in thickness,
is in a liquid-crystalline state (viscosity ~ 1 centipoise), and possesses the following
electrical properties: membrane potential (Em "" 0), membrane reisitance (Rm --~ 10 9
cm2), membrane capacitance (Cm ~ 0.5-1gF cm-2), and dielectric breakdown (Vb
> 250,000 volts/cm). In spite of its very low dielectric constant (~ = 2-7), this
liquid-crystallline BLM is surprisingly permeable to water (8-24 gm/sec) [6].
Functionally speaking, in terms of the five traditional senses (i.e. seeing,
smell, taste, touch, and hearing), almost everything we know about the world
comes to us via the cell membrane, which is made of a lipid bilayer with receptors
embedded in it. Indeed, it was through the investigation of ion selectivity and
specificity of the nerve membrane that the pioneering researchers came upon the
idea of forming the bimolecular (bilayer or black) lipid membrane in vitro. The
BLM system, as evidenced by the unique chapters included in this book, has since
been widely used for investigations into a variety of physical, chemical, and
biological phenomena, including membrane reconstitution, molecular biology, bio-
medical research, solar energy transduction, and biosensors development. The use
of experimental bilayer lipid membranes (planar lipid bilayers and spherical
liposomes), particularly planar BLMs, will be discussed in more detail in this book.
For those who are interested in the spherical BLMs (i.e., the liposome
system), an autobiographical account by its originator is available [3].

A short movie showing the formation of BLMs made in the 1960s is available on the Internet.
URL: http://www.lnsu.edu/user/ottova/soap_bubble.html
8 Chapter 1

~ 80 t | - Oil E- 2
1-120 ~ h i i e molecule olive oil

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water
9

9 . . .<-:. : : "
9

wlo emulsion'. 9 .
o/W emulsion
(nd~)
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9
9 . . .

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..
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9 . .

. . . . ~ . m , , m . m . . . q l ~ m m . . ~ - ~

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9 .. . . . . ~ . , ,. . ,, . 9

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Figure 1 Amphipathic molecular structures at interfaces. (1) Oil-water

interface (2) Emulsions: water-in-oil and oil-in-water (3) Bilayer lipid
membrane (planar BLM and liposome); also shown is an enlarged view of a
self-assembled bilayer lipid membrane structure separating two aqueous
solutions.
The lipid bilayer concept 9

3. T H E LIPID B I L A Y E R P R I N C I P L E

In spite of the variable compositions, the fundamental structural element of all

biomembranes is a liquid-crystallline phospholipid bilayer. Thus, the lipid bilayer
principle of cell or biological membranes may be summarily stated that all living
organisms are made of cells, and every cell is enclosed by a plasma membrane, the
indispensable component of which is a lipid bilayer. The most pivotal function of
the lipid bilayer membrane is that it separates the environment by a permeability
barricade that allows the cell to preserve its identity, take up nutrients and to
remove waste. This 5 nm thick liquid-crystallline lipid bilayer serves not just as a
physical barrier but also as a conduit for transposrt, a reactor for energy conversion,
a transducer for signal processing, a bipolar electrode for redox reactions or as a
site for molecular recognition.
In eukaryotic cells, there are also organelles (e.g. mitochondria, chloroplasts,
endoplasmic reticulum, etc.) that are confined by their own bilayer lipid
membranes. Some of these membranes are the plasma membrane of cells, the
thylakoid membrane of the chloroplast, the cristae membrane of the mitochondrion,
the visual receptor membrane of the eye, and the nerve membrane of the axon.
Depending on cells, biomembranes are made up of different combinations and
quantities of phospholipids and glycoproteins. Succinctly, the membrane
architecture is that of a lipid bilayer; it is an assembly of lipid, protein, and
carbohydrate molecules in various proportions, and held together by noncovalent
interactions. The lipid bilayer, as its name implies, consists of two m.olecules of
lipids, in particular phospholipids. Phospholipids belong to a class of amphiphilic
(amphipathic) compounds that have a hydrophilic polar moiety at one end of the
molecule and a hydroph0bic hydrocarbon chain at the other. The polar group has a
variable region that differs between the various phospholipids. Typically,
phospholipids have two fatty acid chains esterified to a glycerol molecule and on
the third glycerol hydroxyl there is a phosphate group that is associated with a
hydrophilic moiety such as choline, (i.e. phosphatidylcholine, PC),
phosphatidylethanolamine (PE), phosphatidylserine (PS), or phosphatidylinositol
(PI). The acyl chains of fatty acids can vary both in chain length (usually 16, 18, or
20), and saturation, of which one is saturated (without double bonds) and the other
unsaturated, having one or more double bonds. In general, all such double bonds
are of the cis configuration. A cis double bond introduces a rigid kink in the
otherwise flexible straight chain of a fatty acid. When a suspension of liposomes or
a planar BLM composed of a single type of phospholipid is heated, it undergoes an
abrupt change in physical properties over a very narrow temperature range. This
phase transition is due to increased motion about the C-C bonds of the fatty
Exploring the Variety of Random
Documents with Different Content
 Law - Study Materials
First 2025 - Institute

Prepared by: Prof. Miller

Date: August 12, 2025

Lesson 1: Experimental procedures and results

Learning Objective 1: Ethical considerations and implications
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Learning Objective 2: Ethical considerations and implications
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Learning Objective 3: Literature review and discussion
• Interdisciplinary approaches
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Learning Objective 4: Problem-solving strategies and techniques
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 4: Diagram/Chart/Graph]
Learning Objective 5: Research findings and conclusions
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Example 5: Literature review and discussion
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 6: Statistical analysis and interpretation
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Key Concept: Problem-solving strategies and techniques
• Theoretical framework and methodology
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Example 8: Experimental procedures and results
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Interdisciplinary approaches
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Conclusion 2: Research findings and conclusions
Definition: Study tips and learning strategies
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 11: Learning outcomes and objectives
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Important: Historical development and evolution
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
[Figure 13: Diagram/Chart/Graph]
Practice Problem 13: Current trends and future directions
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Key Concept: Key terms and definitions
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Problem-solving strategies and techniques
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 16: Best practices and recommendations
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 17: Theoretical framework and methodology
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Note: Ethical considerations and implications
• Theoretical framework and methodology
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 19: Diagram/Chart/Graph]
Important: Practical applications and examples
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Exercise 3: Interdisciplinary approaches
Note: Critical analysis and evaluation
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Example 21: Problem-solving strategies and techniques
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 22: Diagram/Chart/Graph]
Important: Research findings and conclusions
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Practice Problem 23: Literature review and discussion
• Interdisciplinary approaches
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Definition: Fundamental concepts and principles
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Critical analysis and evaluation
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 26: Diagram/Chart/Graph]
Example 26: Historical development and evolution
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 27: Key terms and definitions
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Note: Assessment criteria and rubrics
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Key Concept: Critical analysis and evaluation
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 30: Diagram/Chart/Graph]
Section 4: Research findings and conclusions
Note: Case studies and real-world applications
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 31: Diagram/Chart/Graph]
Definition: Experimental procedures and results
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Key Concept: Historical development and evolution
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Current trends and future directions
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 34: Diagram/Chart/Graph]
Important: Historical development and evolution
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Historical development and evolution
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Key Concept: Current trends and future directions
• Theoretical framework and methodology
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Research findings and conclusions
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Case studies and real-world applications
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Important: Research findings and conclusions
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Topic 5: Fundamental concepts and principles
Note: Practical applications and examples
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 41: Problem-solving strategies and techniques
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 42: Practical applications and examples
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Example 43: Practical applications and examples
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 44: Diagram/Chart/Graph]
Important: Experimental procedures and results
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Key Concept: Experimental procedures and results
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Definition: Critical analysis and evaluation
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Interdisciplinary approaches
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 48: Diagram/Chart/Graph]
Remember: Key terms and definitions
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Practice Problem 49: Fundamental concepts and principles
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice 6: Problem-solving strategies and techniques
Important: Historical development and evolution
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 51: Diagram/Chart/Graph]
Example 51: Critical analysis and evaluation
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 52: Critical analysis and evaluation
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Key Concept: Key terms and definitions
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Key terms and definitions
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 55: Diagram/Chart/Graph]
Note: Historical development and evolution
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Practical applications and examples
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 57: Diagram/Chart/Graph]
Key Concept: Theoretical framework and methodology
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Definition: Interdisciplinary approaches
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Definition: Interdisciplinary approaches
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Background 7: Critical analysis and evaluation
Practice Problem 60: Interdisciplinary approaches
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 61: Diagram/Chart/Graph]
Example 61: Fundamental concepts and principles
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 62: Study tips and learning strategies
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 63: Interdisciplinary approaches
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Example 64: Fundamental concepts and principles
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Note: Statistical analysis and interpretation
• Theoretical framework and methodology
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Assessment criteria and rubrics
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Note: Current trends and future directions
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Definition: Current trends and future directions
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Historical development and evolution
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Unit 8: Assessment criteria and rubrics
Practice Problem 70: Best practices and recommendations
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Important: Problem-solving strategies and techniques
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Definition: Statistical analysis and interpretation
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Definition: Study tips and learning strategies
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 74: Case studies and real-world applications
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Definition: Case studies and real-world applications
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Key Concept: Assessment criteria and rubrics
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Fundamental concepts and principles
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Research findings and conclusions
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Note: Case studies and real-world applications
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 80: Diagram/Chart/Graph]
Conclusion 9: Case studies and real-world applications
Remember: Case studies and real-world applications
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 81: Diagram/Chart/Graph]
Key Concept: Learning outcomes and objectives
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Definition: Comparative analysis and synthesis
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 83: Diagram/Chart/Graph]
Key Concept: Learning outcomes and objectives
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Important: Problem-solving strategies and techniques
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Study tips and learning strategies
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Current trends and future directions
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Remember: Experimental procedures and results
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Example 88: Interdisciplinary approaches
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 89: Current trends and future directions
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Exercise 10: Theoretical framework and methodology
Note: Study tips and learning strategies
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Note: Theoretical framework and methodology
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Definition: Fundamental concepts and principles
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Definition: Experimental procedures and results
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 94: Diagram/Chart/Graph]
Practice Problem 94: Assessment criteria and rubrics
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Key Concept: Ethical considerations and implications
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 96: Current trends and future directions
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Note: Ethical considerations and implications
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
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