Outline of Lecture 3
file:///C|/Users/dbm2/Documents/COURSES/C2006/current-lectures14/lect3.14.html[1/29/2014 10:18:17 AM]
C2006/F2402 '14 -- Outline Of Lecture #3 -- Last update 01/29/2014 10:10 AM -- Two clarifications were added after the live
lectures. They are in blue.
(c) 2014 Deborah Mowshowitz, Department of Biological Sciences, Columbia University, New York NY
Handouts*: 3A -- Freeze Fracture; Types of Membrane Proteins
3B -- RBC Membrane, RBC -- Role of Anion Exchanger
3C -- ECM (Extracellular Matrix)
Pictures (Power Point Slides) shown at the start of the lecture are on Courseworks for registered students.
The main menu page includes a link to the web-sites page. This page has links to web sites that you may find
interesting and/or helpful. The web sites contain animations, explanations, pictures etc. that are relevant to this course.
(The list is not complete; I'll add to it as we go.) I will add specific links in the lectures, but you may want to explore
some of the sites on your own. Please let me know if any of the web sites are useful, and/or if you find any other good
ones.
I. Introduction to Membrane Structure
A. The Big Question: What does the structure seen in the EM represent? For possibilities, see Becker fig. 7-3. For
an EM picture, see PPt slides, slide #1 or Becker, fig. 7-4.
B. Lipid part
1. Amphipathic nature of lipids -- See Sadava fig. 6.2 -- there are multiple different "two headed" lipids -
- each type has a different structure, but each has a hydrophobic end and hydrophilic end.
2. Amphipathic Lipids form a bilayer.
C. Protein part -- where are the proteins (relative to the lipid)? Is it a "unit membrane" or a "fluid mosaic?"
For "unit membrane" See Becker fig. 7-4 or PPT slide #1 for EM picture; for fluid mosaic model see
Becker fig. 7-5 or Sadava fig. 6.1.
1. Use of freeze fracture procedure
a. E vs P faces of bilayer = surfaces you see if you crack bilayer open = inside of bilayer
(1). E face = inside of the monolayer that is closer to extracellular space (outside
of cell)
(2). P face = inside of the monolayer that is closer to protoplasm (inside of cell)
b. What do you see on inside? See Becker fig. 7-16 & 7-17 or Sadava fig. 6.4 or top panel
on handout 3A.
(1) Inside is not smooth -- shows proteins go through bilayer (implies "mosaic"
model not unit membrane)
(2). More bumps (proteins) on P face than E face -- shows more proteins
anchored on cytoplasmic (protoplasmic) side.
2. Freeze fracture vs Freeze etch
Outline of Lecture 3
file:///C|/Users/dbm2/Documents/COURSES/C2006/current-lectures14/lect3.14.html[1/29/2014 10:18:17 AM]
a. Freeze fracture = crack frozen sample open, examine in EM;
b. Freeze etch = crack open, let some water sublime off to expose deeper layers, then look in
EM. For some sample pictures, see Becker figs. 15-15 (15-16), 15-18 (15-19), 15-24 (15-25),
& 16-1.
D. Fluid mosaic model -- overview of current idea of how proteins and lipids are arranged. See Becker fig. 7-5 (or
7-3) or Sadava fig. 6.1. Also handout 3A, middle of top panel.
II. Fluid Mosaic Model of Membrane Structure
A. Fluid Part = Lipid bilayer
1. Formation of Bilayer -- All amphipathic lipids form bilayers. In cell, virtually all lipids are inserted
from one side of the bilayer of the ER (side facing the cytoplasm). How do lipids get to the other half of
the bilayer?
2. Lateral diffusion vs. flip-flop of lipids. See Becker 7-10 & 7-11.
lateral diffusion = movement within plane of membrane -- fast (secs). Animation of lateral
diffusion. Also see Sadava fig. 6.5 or Becker fig. 7-28 & 7-29.
flip-flop = movement from one side of bilayer to the other -- slow (hrs) w/o enzymes. Enzymes
(flipases = phospholipid translocators) are needed to speed flip-flop. (More details when we get to
transport.)
3. Two sides of a bilayer often have a different lipid composition. (One side = 1/2 of bilayer = a leaflet.)
B. Mosaic Part = Protein. Types of Membrane Proteins -- what do you get if you take a membrane apart? See
handout 3A, bottom panel.
1. Peripheral membrane proteins vs. integral membrane proteins
Type of
Membrane
Protein
Alt. terminology
Protein Removed
From Membrane By
Location/Attachment of Protein
Peripheral Extrinsic salt, pH changes
On one 1 side of bilayer; non covalently attached
to lipid
Integral Intrinsic disrupting lipid bilayer
Goes through bilayer* or Covalently attached to
lipid on one side (Lipid-anchored)**
* A small number of integral proteins do not go all the way through the membrane; they will be largely ignored in this
course. For examples see Becker fig. 7-19 (first protein on left) or Sadava fig. 6.1 -- last protein on right.
**Note that lipid-anchored proteins can be considered a type of integral protein or a separate category. See Becker fig.
7-19.
2. Transmembrane proteins of plasma membrane (See Sadava fig. 6.3 and/or Becker fig. 7-19 & 7-21)
a. Single pass vs multipass
b. Domains -- intracellular, extracellular, transmembrane. Note: For a multipass protein, each
individual section or stretch of polypeptide (transmembrane, extracellular, or intracellular) is
Outline of Lecture 3
file:///C|/Users/dbm2/Documents/COURSES/C2006/current-lectures14/lect3.14.html[1/29/2014 10:18:17 AM]
usually considered a separate domain. See answer to problem 1-20. All the extracellular or all
the intracellular domains may cluster together, but the term 'domain' is not usually used for
the entire extracellular (or intracellular) part of the protein.
c. Location of carbohydrates -- all in extracellular domain (all added inside EMS)
d. Anchorage -- Some proteins are anchored to cytoskeleton; some float in lipid bilayer.
e. Types & Functions -- All bridge the membrane but function differs. Can be:
(1). Transport proteins -- Allow transport of small molecules in and out of cells.
('pumps' or 'doors'.)
(2). Receptors -- Trap (bind) molecules on outside. Then receptor can facilitate:
(a). Transport -- By invagination of membrane. Trapped large molecules are
transported into cell in a vesicle (by RME -- receptor mediated endocytosis).
Example: Receptor for LDL (low density lipoprotein -- a carrier for cholesterol).
(b). Transmission of signals -- relays signals to inside of cell from trapped
molecule on outside of cell. Example: Receptor for Acetyl Choline (a
neurotransmitter).
(c). Both -- can facilitate both transmission of signal and internalization of
signal molecule. Typical Examples: Receptors for growth factors. Specific
example: Receptor for EGF. (EGF = Epidermal Growth Factor)
(3). Connectors -- physically connect cytoskeleton (inside of cell) to materials on
outside of cell (ECM = extracellular matrix) or to next cell. Example: cadherins
(connect cells) & integrins (connect cells and ECM). More on this next time.
(4). More than one of the above -- some transmembrane proteins act in more
than one capacity. Examples: Integrins (connectors & act in signaling).
III. The Red Blood Cell (RBC) Membrane -- The best studied example of a Membrane. For pictures
of RBC see the PPt slides for lecture #3.
A. Why RBC's
1. Easy to get
2. No internal membranes -- all organelles lost during maturation of human RBC -- see Becker fig. 7-20
(a). Only membrane = plasma membrane.
3. Can make 'ghosts' = resealed plasma membranes. Can be resealed (or broken and reformed into
vesicles) in either orientation -- "right" or "wrong" side out. See PPt slides, slide #4.
B. RBC membrane proteins -- Structure & Function. See Becker fig. 7-20 (b) & 15-19 (15-20). (Handout 3B --
top)
1. Peripheral proteins -- spectrin, ankyrin, (band 4.1), actin. Comprise peripheral cytoskeleton, which
supports membrane. All cells are thought to have a similar structure under the plasma membrane.
2. Intrinsic proteins -- Two basic kinds -- single pass & multipass.
a. Example of RBC single pass -- glycophorin -- function of protein not known.
Outline of Lecture 3
file:///C|/Users/dbm2/Documents/COURSES/C2006/current-lectures14/lect3.14.html[1/29/2014 10:18:17 AM]
(1). Has large amount of (-) charged modified carbohydrate -- sialic acid.
Possible functions:
(a). Neg. charge may cause RBC to repel each other and prevent
clumping of RBC.
(b). Loss of terminal sugars may occur with age and trigger
destruction of "old" RBC.
(2). Glycophorins make up a gene family; variations in glycophorin A are
responsible for MN blood type differences. Variations in glycophorin C are
correlated with resistance to malaria.
b. Example of RBC Multipass -- band 3/anion exchanger -- Catalyzes reversible exchange
of the anions HCO
3
-
(bicarb) and Cl
-
between RBC and plasma. Exchange allows max.
transport of CO
2
in blood (as bicarb in solution). See Sadava fig. 49.14 or Becker 8-3.
(1). Why is transport of CO
2
an issue? Tissues carry out oxidative metabolism
and generate lots of CO
2 .
The
CO
2
diffuses out of the cells into the blood.
However the solubility of CO
2
in plasma
(cell-free liquid portion of the blood) is
limited.
(2). Basic idea: Bicarb is much more soluble in plasma than CO
2
, so lots of
bicarb (but not much CO
2
) can be carried in the blood. Therefore need to covert
CO
2
to bicarb when want to carry CO
2
in blood; need to do reverse to eliminate
the CO
2
(in lungs).
(3). Role of Carbonic anhydrase: Conversion of CO
2
to bicarb (& vice versa)
can only occur inside RBC, where the enzyme carbonic anhydrase is. (See
handout 3B, middle panel.) Carbonic anhydrase catalyzes:
CO
2
+ H
2
O HCO
3
-
+ H
+
(4). Role of exchanger: Gases can pass through membranes by diffusion -- CO
2
can exit or enter RBC as needed. However bicarb cannot pass through
membranes. You need the anion exchanger to get bicarb in and out of RBC.
(5). Physiological Function of Exchanger
(a). Where CO
2
is high, as in tissues, CO
2
diffuses into RBC and is
converted to bicarb inside the RBC. (Reaction above goes to right.)
Then bicarb leaves RBC in exchange for chloride using the anion
exchanger.
(b). In lungs, the process is reversed -- bicarb reenters the RBC in
exchange for chloride using the anion exchanger. The bicarb is
converted back to CO
2
inside the RBC (reaction above goes to left).
Then the CO
2
diffuses out of the cells and is exhaled.
(c). To be sure you have this straight, fill in the spaces in the table
Outline of Lecture 3
file:///C|/Users/dbm2/Documents/COURSES/C2006/current-lectures14/lect3.14.html[1/29/2014 10:18:17 AM]
below with 'in' or 'out'. For RBC in each location, does the substance
(CO
2
, bicarb or Cl
-
) go in to the RBC or out of the RBC?
Note: In the table, fill in what happens in the RBC, not what happens
in the tissue or lung cells. Also note that RBC are always in the
blood, inside blood vessels. 'RBC in the lungs' means 'the RBC inside
the blood vessels that pass through the lungs.'
Location of RBC
Lungs Tissues
CO
2
Bicarb
Cl
-
(6). Note on structure & terminology -- in picture on handout, anion exchanger
looks like a channel allowing simple diffusion of bicarb and Cl
-
in and out. (It is
called a channel in some earlier literature.) Exchanger is actually more complex -
- has moving parts and movement of each ion depends on movement of the other.
More details on this & other types of transport proteins next time.
C. Proteins of Other Membranes -- Membranes of other cells are similar. In other membranes:
Find proteins of same protein families as in RBC, as well as entirely different proteins.
Find both intrinsic and peripheral proteins
Intrinsic/integral proteins are both single and multipass proteins; anchored and floating.
Different membrane proteins are found in different cell types.
Try problems 1-2 & 1-3. To review membrane structure, try 1-15 to 1-17 & 1-20.
IV. Extracellular Matrix (ECM) See handout 3C or PPt slide #5.
Note: Becker Chap. 17 goes well beyond what will be covered in this section. References to pictures and diagrams are
included FYI. For the picture in ppt slide #5 go to http://wiki.pingry.org/u/ap-biology/images/5/52/Image122.gif. The
dark purple 'worms' in the picture are adhesive proteins, such as fibronectin. (Alternatively, search Google images for
extracellular matrix. You will see several versions of this picture.)
A. Where do components of the ECM come from? All these components are made inside the cells, and then
secreted -- details of secretion later.
B. What are the major components of the ECM (of animal cells)? Listed on handout 3C, top panel. For a picture
of the ECM, see the bottom panel.
1. Structural proteins. Major ones are
collagen -- nice picture in Becker fig. 17-14 (17-13)
elastin -- diagram in Becker fig. 17-16 (17-15).
2. Adhesive Glyco-Proteins -- fibronectins, laminins, etc. Have multiple binding domains. Connect other
materials in ECM with each other and/or connect to extracellular domains of transmembrane proteins. For structures
Outline of Lecture 3
file:///C|/Users/dbm2/Documents/COURSES/C2006/current-lectures14/lect3.14.html[1/29/2014 10:18:17 AM]
see Becker figs. 17-18 (17-17) for fibronectin & 17-21 (17-20) for laminin. For role of fibronectin in guiding
migrating cells, see Becker fig. 17-19 (17-18).
3. Proteoglycans -- special type of glycoprotein consisting of lots of carbohydrate attached to a protein core.
Provides a gel-like matrix for ECM. See Becker fig. 17-17 (17-16) or handout 3C, bottom panel. Table below is for
reference purposes only so you can follow the terminology.
See http://themedicalbiochemistrypage.org/glycans.html for a nice web site with a summary of structure, function, and
medical significance of proteoglycans and GAGs.
Proteoglycan (mucoprotein) Glycoprotein
General
description
Lots of carbohydrate attached to a
protein core;* Can be 95% carbohydrate
A protein with some carbohydrate
attached
Are sugar
chains
branched?
No Yes
Length of
Sugar Chain
Long Short
Are sugars
repeating?
Yes (repeating disaccharide); & sugars
usually modified
No
Name of
Carbohydrate
Mucopolysaccharide or GAG#
(glycosoaminoglycan)
Oligosaccharide
Example(s)# See Becker fig. 17-17 (17-16). Band 3 protein or glycophorin
Location Extracellular matrix (form gel);
important in knees and other joints.
Integral membrane protein
(carbohydrates on extracellular
domain)
* Multiple proteoglycans can be attached to a core carbohydrate chain (GAG or mucopolysaccharide) to form a giant
aggregate as shown on handout 3C or in Sadava fig. 5-22 (5-25) or Becker fig. 17-17 (17-16) or http://www.in-vivo-
health.co.uk/image/Proteoglycan%20Aggregate2.jpg
#Name of GAG depends on the sugars in the chain. See figure 17-17 in Becker. Examples:
Heparin -- Widely used as an anticoagulant. Inhibits factor required for blood clotting. (Physiological role, meaning
real job in body, may or may not involve inhibition of blood clotting.) Similar to heparan sulfate shown in 17-17.
Chrondroitin sulfate -- Often recommended as a dietary supplement (plus glucosamine) in treatment of arthritis.
Recent results indicate it may be helpful in a small group of patients but is not a panacea.
C. Connection of ECM to cytoskeleton -- ECM often connected to transmembrane proteins called integrins.
Integrins link ECM and cytoskeleton. More details below & next time.
D. Basal Lamina -- see Becker 17-20 (17-19) -- important part of ECM
1. Structure -- Solid layer found in parts of ECM. Main components are networks of laminin & collagen.
For structure of laminin, see Becker fig. 17-21 (17-20)
2. Location -- surrounds some cells (skeletal muscle, fat) and underlies some epithelial layers (on basal or
body side). More details of epithelia next time.
3. Terminology -- Also called basement membrane especially in older literature. Has no lipid & is not a
real membrane.
Outline of Lecture 3
file:///C|/Users/dbm2/Documents/COURSES/C2006/current-lectures14/lect3.14.html[1/29/2014 10:18:17 AM]
4. Function -- physical barrier, support and/or filter.
5. How Connected to cells -- through integrins.
Next time: Cell-Cell (& Cell-ECM) Connective Structures Then, what does a real cell look like? Where are the
connective structures?
