BIOCHEMISTRY OF THE

MUSCLE
BY

PROF MRS J E
IKEKPEAZU
 BIOCHEMISTRY OF THE
MUSCLE
• The muscle is an aggregate of proteins
involved in contraction.
• The musculature(a system of muscles in
the body eg the biceps and triceps in the
arm are part of the musculature of the
arm) is what makes movement possible.
• Muscle is a major biochemical transducer
• Converts potential (chemical) energy into
kinetic (mechanical) energy.
• Muscle •
• They comprise the largest group of tissues in
body
There are 3 types of muscle
• Skeletal muscle: Make up muscular system
• They are the ones that make up
musculoskeletal system
• Cardiac muscle: Found only in the heart
• Smooth muscle: Appears throughout the
body systems as components of hollow organs
and tubes
• Classified as: Striated/unstriated and
voluntary/involuntary (see below)
 CLASSIFICATION OF MUSCLES
They are classified into 3 basic types:
• Skeletal muscle
• Cardiac muscle
• Smooth muscle
• They are also divided into 2 based on
electron microscopic appearance into
• Striated muscles eg cardiac and
skeletal
• Non striated muscles eg smooth
muscles
 Classification continued

• Divided based on control from

the CNS into
• Voluntary muscles eg
skeletal muscles
• Involuntary muscles eg
cardiac and smooth muscles.
• Muscle Action
The controlled muscle contraction
allows :
• Purposeful movement of the
whole body or parts of the body
• Manipulation of external objects
• Propulsion of contents through
various hollow internal organs
• Emptying of contents of certain
organs to external environment eg
• In all of these types of muscle,
contraction is based on the
interplay between two kinds
of protein filaments:
• actin (called the thin
filament) and
• myosin (called the thick
filament).
 ORGANISATION OF THE SKELETAL MUSCLE

• The skeletal
muscle is
striated and
consists of
parallel
bundles of
muscle fibers
connected to
tendons at
both ends.
• Structure of Skeletal Muscle
• Muscle consists of a number of
muscle fibers lying parallel to one
another and held together by
connective tissue
• Single skeletal muscle cell is known
as a muscle fiber
• Multinucleated
• Large, elongated, and cylindrically
shaped
• Fibers usually extend entire length of
• Each muscle fiber is composed of
several myofibrils arranged in
parallel.
• Myofibrils are surrounded by an
electrically excitable membrane
called the sarcolemma.
• The myofibrils are immersed in a
cytosol (Sarcoplasm).
• Sarcoplasm is rich in glycogen, ATP,
creatine phosphate and
glycolytic enzymes.
• Myofibrils exhibit a longitudinally long
repeating structure called the sarcomere,
which is the functional unit of a myofibril.
• Sarcolemmal invaginations gives rise to
small membranous folds, known as the
transverse tubules (T-tubules) which extend
from the sarcolemma and surround each
myofibril at the Z line-see later.
• The sarcoplasmic reticulum is a sheath of
flattened vessicles that surround the
myofibrils (like a net) and stores large
quantities of calcium at its terminal cisternae
.
 Electron micrograph of the
sarcomere
• The sarcomere is characterized by the
appearance of several distinct bands.
• The less optically dense band is referred to
as the I band, and the denser one as the A
band.
• A dense line appears in the centre of the 1
band, called the Z line or disc, and a dense
narrow band, somewhat similar in
appearance also occurs in the centre of A
band, called the M line.
• Adjacent to the M line are regions of the A
band that appear lense dense than the
• Transverse sections of the
sarcomere reveal that the above
pattern result from the
interdigitation of the two sets of
protein filament- the thin (ACTIN)
and the thick (MYOSIN) filaments.
• The 1 band consists of the thin
filaments, while the H zone
consists of thick filament but the A
band shows a regularly packed
array of interdigitating thick and
thin filaments-see below.
• Organizational details of a typical striated skeletal
muscle. a: Representation of each muscle fiber
showing the parallel bundles called myofibrils.
• b: Myofibrils are a series of sarcomeres separated by Z
discs or lines which contain thick and thin filaments.
• c: Thick filaments are myosin bundles that span the A
band and are bound to proteins of the M line and to
the Z discs across the I bands by the large protein
called titin-see below.
• d: Transmission electron micrograph (TEM) showing
the molecular organization of the sarcomere. e: TEM
of an oblique section showing the hexagonal
organization between the thick and thin myofilaments.
• The organization of individual contractile proteins
making up a sarcomere is a key feature of the
sliding filament model.
• Each sarcomere is composed of hundreds of
filamentous protein aggregates, each known as a
myofilament.
• Two kinds of myofilaments are identifiable on the
basis of their diameter and protein composition
(see image above).
• Thick myofilaments are composed of several
hundred molecules of one of several different
fibrous proteins known as myosin.
• Thin myofilaments are composed of two helically
interwound, linear polymers of globular proteins known
as actin.
• Thin and thick filaments also contain accessory proteins
as described below.
• Proteins of the Z disc, primarily the α-actinins (see later),
serve as an embedding matrix or anchor for one end of
the thin filaments, which extend toward the center of
sarcomeres on either side of the Z disc.
• The Z disc proteins often appear continuous across the
width of a muscle fiber and seem to act to keep the
myofibrils within a myofiber.
• Proteins of the myofibril –
• Thin and thick filaments.
The thin filaments are composed of
three proteins
• Actin
• Troponin.
• Tropomyosin
• Actin is the major constituent
(protein) of thin filaments.
• Comprises 25% of muscle protein by
weight.
• The monomer of Actin is called the G-
actin (because of its globular shape).
• G-actin polymerizes as ionic strength
increase to physiological level into a
fibrous form called F-actin.
• Each actin filament (F-actin) consists
of two stands of actin twisted into an
-helical pattern.
• An F-actin fibre looks
(micrographycally) like two strings of
beads wound around each other in
an  helical form.
Polymerization of G-Actin to F-Actin
Sequence of polymerization of G-Actin
FORMATION AND ASSEMBLY OF THE THIN FILAMENT
F-Actin showing exposed binding site usually covered by tropomyosin- role of calcium ions.
• Tropomyosin (TM) is a
filamentous protein containing
two subunits wound in an -
helical form.
• It lies in the groove on either side
of the F- actin filament
• It mediates access to the site on
actin monomers unto which
myosin head binds.
 Present in cells and muscle-like
structures
• Troponin is a complex of three non-
identical subunits:
• TN-C (a calcium binding subunit),
• TN-T (a TM-binding subunit) and
• TN-1 (an “inhibitory” subunit).
• Two molecules of troponin (TN) bind
to the actin filament at each helical
repeat.
• Troponin T binds to tropomyosin
and together with troponin I, inhibits
the interaction of actin and myosin.
• Troponin I binds actin and inhibits the
binding of actin to myosin ie has a
strong affinity for actin. In other words, it
inhibits F-actin –myosin interactions
• Binds to other troponin molecules
Troponin C
• a calcium binding protein (similar to
calmodulin in function)
• has a binding site for Ca2+
• Binds 4 molecules of calcium
• when calcium is bound, actin and myosin
interaction is promoted.
 NOTE:
• Troponin is found only in
striated muscle cells.
• The thick and thin filaments
interact via cross bridges
• Interaction between this cross
bridges and actin filament cause
contraction
• The filaments slide pass one
another during contraction.
 The thick filaments-MYOSIN

• The thick filaments are composed of

myosin molecules which are long
fibrous structures composed of six
subunits (a hexamer): two heavy
chains and four light chains.
• Contributes 55% of muscle protein by
weight
• Form the thick filament
• Has a fibrous portion consisting of 2
intertwined helices
• Each myosin heavy chain consists of a
globular head and a fibrous tail.
• The two fibrous tails of the heavy chains are
twisted into a double helical structure,
forming regions where myosin molecules
interact to form filaments.
• The globular head of the myosin molecule
contain ATpase activity as well as site for
binding to actin filaments.
• The helical coils therefore form the backbone
of the thick filaments.
• They also form an arm that can provide a
flexible extension or hinge connecting the
globular head to the body of the thick
filament.
DETAILED STRUCTURE OF MYOSIN
 Summary of notes on myosin and actin
Myosin
• Component of thick filament
• Protein molecule consisting of two identical subunits shaped
somewhat like a golf club racket
• Tail ends are intertwined around each other
• Globular heads project out at one end-see structure above.
• Tails oriented toward center of filament and globular heads
protrude outward at regular intervals on each filament
• Heads form cross bridges between thick and thin filaments
• Cross bridge has 2 important sites critical to contractile
process • An actin-binding site • A myosin ATPase (ATP-
splitting) site
• Actin
• Primary structural component of thin
filaments.
• Spherical in shape
• Thin filament also has 2 other proteins
• Tropomyosin
• Troponin
• Each actin molecule has special binding site
for attachment with myosin cross bridge
• Binding results in contraction of muscle fiber
 Summary of notes on myosin and actin CTD
• Actin and Myosin •pls note these: Actin and myosin
are often called contractile proteins.
• Actin and myosin are not unique to muscle cells, but
are more abundant and more highly organized in
muscle cells
• Tropomyosin and Troponin
• Often called regulatory proteins
• Tropomyosin
• Thread-like molecules that lie end to end alongside
groove of actin spiral
• In this position, covers actin binding sites, blocking
interaction of actin and myosin that should lead to
muscle contraction
• Troponin
• Made of 3 polypeptide units
• One binds to tropomyosin(TNT)
• One binds to actin(TNI)
• One can bind with Ca2+(TNC)
• Tropomyosin and Troponin
• Troponin
• When not bound to Ca2+
• Troponin stabilizes tropomyosin in blocking position over
actin’s cross-bridge binding sites
• When Ca2+ binds to troponin
• Tropomyosin moves away from blocking position
• With tropomyosin out of the way, actin and myosin bind,
 ENZYMATIC HYDROLYSIS OF MYOSIN
• Myosin can be cleaved enzymatically into
functional fragments that retain some of
the activities of the intact molecule.
• Myosin is split by trypsin into two
fragments called light meromyosin
(LMM) and heavy meromyosin (HMM).
• LMM like myosin tail forms filament but
lack ATpase activity and does not
combine with actin.
• HMM catalyzes the hydrolysis of ATP and
binds to actin, but does not form
filaments.
• The LMM is a two stranded  helical rod
• They are insoluble alpha-helical fibres
• HMM consists of a short rod attached to
a double headed globular region
• can be split by papain into two globular
fragments (called S1) and one rod-
shaped subfragment (called S2).
• S2 fragment is fibrous
• Each S1 fragment contain an ATpase
active site and a binding site for actin.
• The light chains of myosin are bound to
the S1 fragments.
 GENERAL MECHANISM OF MUSCLE CONTRACTION

• The major biochemical events

occurring during one cycle of muscle
contraction and relaxation can be
represented in the five steps shown
below:
• (1) In the relaxation phase of
muscle contraction, the S-1 head
of myosin hydrolyzes ATP to ADP and
Pi but these products remain bound.
• The resultant ADP-Pi-myosin complex
has been energized and is in high-
energy conformation.
• (2) When contraction of muscle
is stimulated (via events involving
Ca2+, troponin, tropomyosin, and
actin, which are described below
later), actin becomes accessible.
• The S-1 head of myosin finds it and
binds to it
• forms the actin-myosin-ADP-Pi
complex – so called actinomyosin-
ADP-Pi complex.
• (3) Formation of this complex
promotes the release of Pi,
which initiates the power stroke.
• This step which is followed by release
of ADP is accompanied by a large
conformational change in the head of
myosin in relation to its tail , pulling
actin about 10 nm toward the centre
of the sarcomere.
• This is the power stroke.
• The myosin is now in a low-energy
state, indicated as actin-myosin.
• (4) Another molecule of ATP
binds to the S-1 head, forming
an actin-myosin-ATP complex.
• (5) Myosin-ATP has a low
affinity for actin, and actin is
thus released.
• This last step is a key
component of relaxation and
is dependent upon the binding of
ATP to the actin-myosin complex.
 DETAILS
• STEPS IN MUSCLE CONTRACTION
• sliding filament model of muscle
contraction.
• 1. The process of muscular contraction
entails a sliding of the thick and thin filaments
past each other.
• The movement of myosin (thick filaments)
along the actin ( thin filament) is ATP
dependent.
• This contraction is a cyclical process.
• 2. The myosin head binds ATP, which is
hydrolyzed to ADP and Pi by myosin ATpase.
• ADP and Pi remain bound to the ATpase site.
• 3. The myosin head moves to an
adjacent thin (actin) filament (the
binding site) and as it makes contact
with the actin binding site, Pi is
released.
• The release of pi is the rate-limiting
step in muscular contraction.
• 4. Strong cross-bridges form between
actin and myosin which is followed by a
structural alteration (conformational
change) in the myosin molecules and
an effective translocation of the thick
filament relative to the thin filament.
• ie large conformational change in the
head of myosin in relation to its tail ,
pulling actin about 10 nm toward the
center of the sarcomere.
• During this process, the ADP is released.
• 5. After the translocation step, the
bridge structure is broken by the binding
of ATP, which is hydrolysed to ADP and
pi .
• Hydrolysis of ATP result in the myosin
head resuming its original conformation
and is then ready for another cycle
further along.
• The contraction process therefore
involves the breakage and
reformation of cross bridges between
the actin and myosin molecules in a
reaction that requires the
expenditure of ATP.
• Each thick filament has about
500 myosin heads and each head
cycles about five times per
second in the course of a rapid
contraction.
• In a fully contracted myofibril, the actin
and myosin filament show a maximum
overlap with each.
• However there is no change in the
length of either types of filament
(see diagrams below). In other words
during contraction
• 1. The Z lines approach one
another
• 2. The sarcomere get smaller and
• 3. The A band, does not change
size.
• Note: In a relaxed muscle, the actin
filament extend approximately halfway
over the myosin filament-see above
diagram.
• During contraction, the actin
filaments slide towards each other,
past the myosin filaments resulting
in the shortening of the sarcomere,
the myofibrils and the length of the
muscle.
• This is known as the sliding filament
model of muscle contraction.
• The sliding filament cross-bridge
model is the foundation of current
thinking about muscle contraction.
• The basis of this model is that the
interdigitating filaments slide past one
another during contraction and cross-
bridges between myosin and actin
generate and sustain the tension.
• The hydrolysis of ATP is therefore used to
drive movement of the filaments.
• REGULATION OF MUSCLE CONTRACTION:
ROLE OF TROPONIN AND TROPOMYOSIN IN
MUSCULAR CONTRACTION (ACTIN –BASED
REGULATION).
• This takes place manly in striated muscles
(skeletal and cardiac)
• Control of muscle contraction is provided by
troponin, tropomyosin, and a change in intracellular
calcium concentration.
• When the calcium ion concentration is low (10-
7
M), troponin I and troponin T are strongly bound to
tropomyosin, holding it in the actin groove so that
it blocks/covers the myosin binding site on the
actin monomers.
• Troponin C is either loosely associated with
tropomyosin or is bound to troponin I and troponin
• When calcium concentration increase
above 10-7M in the cell, troponin C binds
calcium and undergoes change in
conformation, forcing troponin I and
troponin T to move.
• The movement exposes the myosin
binding site on the actin by the removal
of tropomyosin, permitting interaction
between the two filaments to occur.
• This however does not necessarily result
in contraction; contraction can result only
if ATP is present.
• CONTROL OF INTRACELLULAR CALCIUM
CONCENTRATIONS.
• Small membranous folds, known as T-
tubules extend from the sarcolemma and
surround each myofibril at the Z line-see
earlier.
• The T-tubules act as a communication
system for the depolarization of the plasma
membrane that occurs when a nervous
stimulation signals the muscle to contract.
• Depolarization of the sarcolemma is relayed
to the sarcoplasmic reticulum (SR), a
sheath of flattened vesicles that surround
the myofibrils and store large quantities of
calcium at the terminal cisterna.
• Normally, extracellular calcium
concentrations are high (10-3M) whereas
those in the cytosol are low (10-7M).
• Upon depolarization, the calcium level
rise from 10-7 to greater than 10-5m
during contraction.
• The increase is due primarily to the
movement of calcium ions through ca2+
release channels in the membrane of
the sarcoplasmic reticulum.
• The change in calcium concentration
occurs rapidly and relieves the
inhibition of myosin binding to actin.
• Once the stimulation of the
nerves cease, calcium is pumped
back into the sarcoplasmic
reticulum by the action of a SR-
specific ATpase .
• The calcium is bound in the SR
calsequesterin, an acidic protein
with a high density of aspartate
and glutamate residues.
• Sequence of events in contraction and
relaxation of skeletal muscle (starting
with the events at the neuromuscular
junction).
• Steps in contraction
• (1) Discharge of motor neuron
• (2) Release of transmitter (acetylcholine) at
motor endplate
• (3) Binding of acetylcholine to nicotinic
acetylcholine receptors
• (4) Increased Na+ and K+ conductance in
endplate membrane
• (5) Generation of endplate potential
• (6) Generation of action potential in muscle
fibers
• (7) Inward spread of depolarization
along T tubules
• (8) Release of Ca2+ from terminal
cisternae of sarcoplasmic reticulum and
diffusion to thick and thin filaments
• (9) Binding of Ca2+ to troponin C,
uncovering myosin binding sites of actin
• (10) Formation of cross-linkages
between actin and myosin and sliding of
thin on thick filaments, producing
shortening
• Steps in relaxation
• (1) Ca2+ pumped back into
sarcoplasmic reticulum
• (2) Release of Ca2+ from troponin
• (3) Cessation of interaction
between actin and myosin

• The details of the other events

between actin and myosin are
already described.
• ACTIN-BINDING DRUGS
• Can interfere with the polymerization-
depolymerization cycle of microfilaments.

• Cytochalasin B inhibits the assembly of actin

filaments by binding to one end of the filament
and preventing the addition of actin molecules
to the filament.
• Processes such as endocytosis, cytokinesis,
cytoplasmic and amoeboid movements are all
inhibited by cytochalasin B.

• Phallodin binds along the length of actin

filament, preventing depolymerization.
ACCESSORY PROTEINS OF
THE MYOFIBRIL.
• ACCESSORY PROTEINS OF THE MYOFIBRIL.
• Several other proteins are associated with the
sarcomere. Their location and functions are as follows:
• Titin - summary
• Giant, highly elastic protein (largest in body)
• Extends in both directions from M line along length of
thick filament to Z lines at opposite ends of sarcomere
• 2 important roles:
• Helps stabilize position of thick filaments in relation to
thin filaments
• Greatly augments muscle’s elasticity by acting like a
spring
• It plays a role in relaxation of muscle.
• Nebulin
• is a large fibrous protein attached to
the Z-line and extends the length of
the thin filament.
• It stabilizes the highly ordered
structure of the myofibril by
regulating the assembly of the actin
filaments.
• α-Actinin
• Anchors actin to Z lines.
• Stabilizes actin filaments.
• Desmin
• is found in Z line, where it holds myofibril in
place.
• It gives the muscle its “Striated” appearance.
• Attaches to plasma membrane (sarcolemma).
• Myosin- binding protein C
• Arranged transversely in sarcomere A-bands
• Binds myosin and titin
• Plays a role in maintaining the structural
integrity of the sarcomere.
• Myomesin- cross-links adjacent filaments at
the M-line (the centre of the sarcomere).
• Calcineurin
• Present in the Cytosol (sarcoplasm)
• A calmodulin-regulated protein
phosphatase.
• May play important roles in cardiac
hypertrophy and in regulating amounts of
slow and fast twitch muscles.
• Dystrophin
• Attached to sarcolemma through
dystroglycans
• Deficient in Duchenne muscular dystrophy.
• Mutations of its gene can also cause dilated
cardiomyopathy.
• The Dystrophin Complex
• The dystrophin-glycoprotein complex (DGC) is a multi-
subunit complex within and across the membranes of
cardiac and skeletal muscle cells as well as vascular
smooth muscle cells.
• The complex serves both a mechanical stabilizing and
a signaling role in these various cell types.
• The complex mediates interactions between the
cytoskeleton, membrane, and the extracellular matrix.
• The complex is composed of up to 15 different
proteins dependent upon its location.
• These proteins include, extracellular , transmembrane,
and intracellular subunits.
• Within the DGC there is the protein dystrophin,
two dystroglycan proteins (α-dystroglycan, α-DG
and β-dystroglycan, β-DG), the syntrophins (α and
β), the dystrobrevins (α and β), caveolin-3,
sarcospan, and a sub-complex composed of
members of the sarcoglycan family.
• There are six known sarcoglycans designated α-, β-,
γ-, δ-, ε (epsilon)-, and ζ (zeta)-sarcoglycan.
• An additional component of the DGC is neuronal
nitric oxide synthase (nNOS, NOS-1).
• The major intracellular protein is the large
dystrophin protein.
Organizational details of a typical dystrophin-glycoprotein complex.
• The dystrophin proteins serves to link the
extracellular portions of the DGC with the actin
filaments within the cell.
• Dystrophin physically interacts with the
cytoplasmic tail of β-DG, via a C-terminal domain
in dystrophin, and γ-actin in actin filaments (F-
actin) via an N-terminal domain.
• The clinical significance of the dystrophin protein
relates to the fact that mutations in the DMD gene
are the cause of various muscular dystrophies (see
below): Duchenne muscular dystrophy (DMD),
Becker muscular dystrophy (BMD), and X-linked
dilated cardiomyopathy (XLDCM). See later
 CONTRACTION OF SMOOTH MUSCLES
• Smooth muscle differs from skeletal
muscle in various ways:
• Smooth muscles are found, for
example, in blood vessel walls and in
the walls of the intestine.
• In smooth-muscle cells, which are
usually spindle-shaped, the contractile
proteins are arranged in a less regular
pattern than in striated muscle.
• The smooth muscle contain actin
filament attached to the plasma
membrane, but the fibrils are not
aligned, thus when myosin slides
along the actin filaments, the cell
contracts in all direction.
• Contraction in this type of muscle is
usually not stimulated by nerve impulses,
but occurs in a largely spontaneous way.
• Ca2+ (in the form of Ca2+-calmodulin);
also activates contraction in smooth
muscle.
• In this case, however, it does not bind to
troponin, but activates a protein kinase
that phosphorylates the light chains in
myosin and thereby increase myosin’s
ATPase activity.
• Most of the Ca2+ are from the ECF
and there is no troponin.
 Sources of calcium ions for
smooth muscle contraction
• The phosphorylation of membrane-bound PI
produces phosphatidylinositol 4,5-
bisphosphate (PIP2).
• This compound is degraded by phospholipase C
β in response to the binding of various
neurotransmitters, hormones, and growth
factors to membrane G protein– coupled
receptors.
• The products of this degradation, inositol 1,4,5-
trisphosphate (IP3) and DAG, mediate the
mobilization of intracellular calcium from the
sarcoplasmic reticulum.
• There is also opening of the calcium ion
channels at the cell membrane that allows
inflow of ca2+ from the ECF.
• Smooth Muscle contraction - Details
• While the sliding filament model
adequately describes the basic mechanism
of contraction in all muscle types, there are
significant differences between striated
(skeletal and cardiac) and smooth muscle.
• Although smooth muscle lacks the troponin
complex, its contractile activity is still
regulated by cytoplasmic calcium levels.
• A Ca2+/calmodulin (CaCM) binding protein known
as caldesmon regulates the movement of
smooth muscle tropomyosin on and off the
myosin binding sites of thin filaments.
• Alterations in smooth muscle cytosolic calcium
levels occur via:
• 1. voltage-dependent activation processes and
by
• 2. receptor-mediated processes.
• The voltage-mediated processes involve the
activation of plasma membrane voltage-gated
calcium channels of the L-type- see above.
• The principal receptor-mediated activation of
smooth muscle contractile activity occurs in
response to α1-adrenergic receptor activation.
• The α1 adrenergic receptor is coupled to a Gq-type
G-protein which activates PLCβ leading to
production of inositol-1,4,5-trisphosphate (IP3)
and diacylglycerol, DAG from P1P2.
• The IP3 binds to receptors on the endoplasmic
reticulum leading to release of stored Ca2+, the
consequences of which are smooth muscle
contraction.
• In contrast to most of the vasculature, the smooth
muscle cells of the vessels in skeletal muscle
tissues possess predominantly the β 2 adrenergic
receptor.
• The β2 adrenergic receptor is coupled to a G s type
G-protein, the activation of which results in
increased cAMP production and activation of PKA.
• Both PKA and cAMP interfere with smooth muscle
contraction leading, instead, to relaxation of the
vessels.
• This allows skeletal muscle cells access to increased
nutrients and oxygen in response to stress.
• Both the voltage-mediated and the receptor-mediated
smooth muscle activation processes lead to increased
intracellular calcium levels which lead to increased levels
of active CaCM which, in turn, binds caldesmon, removing
it from its site on thin filaments.
• Calmodulin is also a component of the myosin light-chain
kinases (MLCK or MYLK) and the activation of calmodulin
(CM or CaM) in these enzyme complexes results in
phosphorylation of myosin light-chains (MLC or MYL) on
serine 19 (S19).
• Concurrently, tropomyosin is observed to change its
location in the helical grooves of F-actin, and the ATPase
activity of the actin-myosin complex (actomyosin) is
stimulated.
• Each of these events results in contraction of smooth
• Myosin Light Chain Kinases
• When calcium is depleted, the CaCM complex
dissociates and caldesmon is released from its
complex with calmodulin and re-associates with
thin filaments while simultaneously, MLCK (MYLK)
activity is reduced.
• The actomyosin complex ATPase activity is
correspondingly inhibited.
• In essence, caldesmon in smooth muscle cells
replaces the troponin complex as a calcium
(CaCM)-dependent regulator of the location of the
tropomyosin complex within the context of the
thin filaments.
• Smooth muscle relaxation is effected via activation
of β2-adrenergic receptors which are coupled to Gs-
type G-proteins.
• Activation of β2-adrenergic receptors leads to
increased production of cAMP via activation of
adenylate cyclase.
• The increased cAMP levels activate PKA which in
turn phosphorylates MLCK (MYLK) resulting in
inhibition of MLCK-mediated phosphorylation of
myosin light chains.
• In addition, cAMP itself will inhibit the activity of
MLCK (MYLK).
• Another path to smooth muscle relaxation initiated
via β2 receptor activation is the PKA-mediated
phosphorylation of a membrane potassium channel
(KATP) which results in depolarization of the cell and
closure of the plasma membrane Ca2+ channels.
• Closure of the Ca2+ channel results in reduced
intracellular Ca2+ and thus, reduced levels of active
MLCK (MYLK).
• Activation of α2-adrenergic receptors can interfere
with the effects of β2 receptor activation since α2-
adrenergic receptors are coupled to Gi-type G-
proteins that inhibit the activity of adenylate
cyclase.
• Myosin Light Chain Phosphatases
• Following removal of the initiating stimulus for
smooth muscle contraction, all of the events that
participated in the activation of contraction must
be reversed.
• With respect to the myosin proteins this entails
dephosphorylation of S19 of the myosin light
chains. Phosphate removal from myosin light
chains is catalyzed by an enzyme called simply,
myosin phosphatase.
• The catalytic activity of myosin phosphatase is a
protein that is a member of the protein
phosphatase 1 (PP1) family of phosphatases.
• Adrenergic Receptors in Muscle Functions
• The catecholamines, norepinephrine and
epinephrine, exert potent effects on cardiac,
skeletal, and smooth muscle cells.
• These effects are exerted in response to binding to
one or more of the different types of adrenergic
receptors – Read up these receptors types
(α1,α2,β1,β2,β3), their expression profile and
functions.
• Acetylcholine and Receptors in Muscle Functions
• Also read up acetylcholine and its receptors and
their role in muscle functions.
Receptor Type Gene Symbol(s) Expression Profile Functions / Comments

three subtypes: α1A, α1B, α1D; coupled to Gq-type G-proteins,

ADRA1A vasoconstrictor for coronary arteries and veins, decreases GI
predominates in heart, blood vessels, and
α1 ADRA1B smooth muscle cell motility, induces contraction of smooth
kidneys, also expressed in adipose tissue
ADRA1D muscle in uterus, urethral sphincter, vas deferens, and ureter,
modulates glycolysis and gluconeogenesis

three subtypes: α2A, α2B, α2C; coupled to Gi-type G-proteins, acts

within the CNS to decrease blood pressure and exert
bradycardic effects, exerts a hypothermic effect, arterial and
ADRA2A
central nervous system (widely distributed); venous vasoconstriction, inhibits insulin release and stimulates
α2 ADRA2B
vessels, adipose tissue, kidneys, and platelets glucagon secretion, modulates gluconeogenesis and glycolysis,
ADRA2C
inhibits gastric acid secretion and gastric motility, inhibits
release of norepinephrine and acetylcholine, involved in
thrombus stabilization by inducing platelet aggregation
 coupled to Gs-type G-protein, exerts inotropic
(contraction strength) and chronotropic (heart rate)
heart, kidney, skeletal muscle, lung,
β1 effects on the heart, increases fat mobilization from
ADRB1 colon, liver, thyroid gland, adipocytes
adipose tissue, increases renin release from kidneys,
(preadipocytes only in BAT)
enhances sensation of hunger through release of ghrelin
by the stomach

coupled to Gs-type G-protein, bronchodilator and

adipose tissue but not brown adipocytes,
vasodepressor, induces relaxation of smooth muscle in
bronchioles, skeletal muscle, smooth
β2 ADRB2 bronchus, bronchioles, uterus, and detrusor muscle, inhibits
muscle, lung, kidney, colon, liver, thyroid
release of insulin, stimulates lipolysis, glycolysis, and
gland, heart
gluconeogenesis

abundant in adipocytes of BAT and

omental fat, gallbladder and bladder, is coupled to Gs-type G-protein, regulation of lipolysis,
β3 ADRB3 not expressed in the heart, skeletal principal norepinephrine receptor in BAT, increase lipolysis
muscle, liver, kidneys, lung, or thyroid in BAT and plays major role in adaptive thermogenesis
gland
• NEUROMUSCULAR DISORDERS
• 1. Myasthenia Gravis (MG)
• This is a particularly devastating disease that results
from defects in the overall processes of neuromuscular
nerve transmission.
• MG is a very serious disorder that is often times fatal.
• The characteristic features of the disease are weakened
skeletal muscles that tire with very little exertion.
• MG is an auto-immune disease associated with
antibodies to the nAChR of the neuromuscular junction.
• Binding of the antibodies to the receptor results in
receptor destruction as well as receptor cross-linking.
• In most patients with MG there is a 70%–90%
reduction in motor end plate nicotinic receptor number.
• Two major forms of MG exist, one in which the
extraocular muscles are the ones primarily affected
and in the other form there is a generalized skeletal
muscle involvement.
• In the latter form of MG, the muscles of the
diaphragm become affected resulting in respiratory
failure which contributes to the mortality of MG.
• Treatment of MG involves numerous approaches
including the use of acetylcholinesterase inhibitors.
• The use of these types of drugs allows for enhanced
levels of ACh at the motor end plate during repeated
• 2. The Muscular Dystrophies
• The muscular dystrophies represent a group of nine
characterized disorders, all of which are associated
with some level of loss of muscle function along
with atrophy of muscle tissue.
• These diseases are Duchenne muscular dystrophy
(DMD), Becker muscular dystrophy (BMD),
myotonic dystrophy (DM, for dystrophia
myotonica), distal muscular dystrophy, Emery-
Dreifuss muscular dystrophy, limb-girdle muscular
dystrophy, oculopharyngeal muscular dystrophy,
fascioscapulohumeral muscular dystrophy, and
congenital muscular dystrophy.
• The muscular dystrophies that are associated with defects in
the gene encoding the intracellular protein, dystrophin (gene
symbol, DMD), are known as Duchenne muscular dystrophy
(DMD) and Becker muscular dystrophy (BMD).
• Because the dystrophin gene is found on the X chromosome,
both of these diseases are inherited in an X-linked manner.
• Both DMD and BMD are caused by mutations in the DMD
gene.
• The primary clinical differences between DMD and BMD are
due to the fact that the mutations in the DMD gene that cause
Duchene muscular dystrophy result in virtually no functional
dystrophin protein being made.
• With Becker muscular dystrophy the mutations result in some
functional dystrophin protein ranging from 10%–40% of
normal.
• Duchenne Muscular Dystrophy, DMD
• Duchenne muscular dystrophy (DMD) represents
the most severe form of nine characterized
muscular dystrophies.
• DMD is an X-linked recessive disorder and,
therefore, primarily manifests in males.
• The disease is inherited with a frequency of
approximately 1 in 3,600.
• DMD is a rapidly progressing, fatal form of
muscular dystrophy.
• Symptoms of DMD usually begin to appear within
the first 6-months of life but can also be seen at
birth in some afflicted infants.
• The symptoms of DMD are characterized by
progressive muscle degeneration and
weakness, eventually resulting in death.
• The average life span for DMD patients is
around 25 years of age.
• The early signs that are characteristic in all
DMD patients is a progressive proximal
muscle weakness evident in the legs and
pelvis.
• These symptoms are associated with, and the
result of, a loss of muscle mass.
• Although muscle loss is characteristic of DMD,
very early in the disease the calf muscles
hypertrophy.
• As the disease progresses the muscle loss and
weakness spreads to the arms, neck, and
other areas of the body.
• As the disease progresses, the loss of muscle
tissue is replaced fatty tissue and fibrotic
tissue.
• Becker Muscular Dystrophy, BMD
• Becker muscular dystrophy (BMD) represents a
milder form of muscular dystrophy caused by
mutations in the dystrophin gene (gene symbol,
DMD).
• Whereas in the case of Duchenne muscular
dystrophy where no functional dystrophin protein
is made, BMD is associated with some functional
protein.
• For this later reason the symptoms of BMD are
much less severe, begin manifesting later in life
than for DMD, and the disorder is not lethal as in
the case of DMD.
• Because BMD is caused by mutations in the same gene,
the location of symptoms is very similar to that in the
case of DMD but just manifest much later and with less
severity.
• Symptoms of BMD usually begin to appear between 5
and 15 years of age.
• Initial symptoms include calf muscle enlargement
followed by the same fatty tissue and fibrotic tissue
deposition as in DMD.
• The muscle weakness in BMD is very slowly progressing
and many afflicted individuals can continue to walk,
though with difficulties, well into adulthood.
• In many cases, patients with BMD die in the 4th decade
of life, but many individuals have also lived a normal life
span.
• MUSCLE METABOLISM 1
• The metabolic profile of the muscle
• The major fuels for muscle are glucose,
fatty acids and Ketone bodies.
• Muscle has a large store of glycogen
(1,200Kcal).
• About 3/4 of all the glycogen in the body is
stored in muscle.
• Muscle contraction is associated with a
high level of ATP consumption.
• Without constant resynthesis, the amount
of ATP available in the resting state would
be used up in less than I second of
contraction.
• A. Energy metabolism in the white and
red muscle fibers
• Muscles contain two types of fibers, the
proportions of which vary from one type of
muscle to another.
• Red fibers (type I or slow fibers) are
suitable for prolonged effort.
• Their metabolism is mainly aerobic and
therefore depends on an adequate supply of
O2.
• White fibers (types 11 or fast fibers) are
better suited for fast, strong contractions.
• These fibers are able to form sufficient ATP
even when there is little O2 available
• With appropriate training, athletes
and sports participants are able to
change the proportions of the two
fiber types in the musculature
• This prepares them for the
physiological demands of their
disciplines in a targeted fashion.
• The expression of functional muscle
proteins can also change during the
course of the training.
• Red fibers provide for their ATP
requirements mainly (but not
exclusively) from fatty acids, which are
broken down via β-oxidation, the
tricarboxylic acid cycle, and the
respiratory chain.
• The red color in these fibers is due to
the monomeric heme protein myogobin
which they use as an O2 reserve.
• Myoglobin has a much higher affinity
for 02 than hemoglobin and therefore
only releases its O2 when there is a
severe drop in O2 partial pressure.
• At a high level of muscular effort-e.g,
during weightlifting or in very fast
contractions such as those carried
out by the eye muscles-the 02 supply
from the blood quickly becomes
inadequate to maintain the aerobic
metabolism.
• White fibers therefore mainly obtain
ATP from anaerobic glycolysis.
They have supplies of glycogen
from which they can quickly release
glucose-1-phosphate when needed.
• By isomerization, this gives rise to
glucose-6-phosphate, the substrate for
glycolysis.
• The NADH+H+ formed during glycolysis
has to be reoxidized into NAD+ in order
to maintain glucose degradation and
thus ATP formation.
• If there is a lack of O2, this is achieved
by the formation of lactate, which is
released into the blood and is
resynthesized into glucose in the liver
(cori cycle) when the cell resumes
aerobic metabolism.
• Muscle-specific auxiliary reactions
for ATP synthesis: These exist in order
to provide additional ATP in case of
emergency.
• [Link] phosphate acts as a buffer
for the ATP level-see below.
• [Link] ATP-supplying reaction is
catalysed by adenylate kinase [1].
This disproportionates two molecules of ADP
into ATP and AMP.
• The AMP is deaminated by AMP deaminase
into IMP in a subsequent reaction [2] in order
to shift the balance of the reversible reaction
[1] in the direction of ATP formation.
• B. creatine metabolism
• Creatine (N-methylguanidoacetic acid) and
its phosphorylated form creatine
phosphate (a guanidophosphate) serve as
an ATP buffer in muscle metabolism.
• In creatine phosphate, the phosphate
residue is at a similarly high chemical
potential as in ATP and is therefore easily
transferred to ADP.
• Conversely when there is an excess of ATP,
creatine phosphate can arise from ATP and
creatine (during the resting phase of
contraction).
• Both processes are catalyzed by creatine
kinase .
• In resting muscle, creatine phosphate
forms due to the high level of ATP.
• If there is a risk of a severe drop in the
ATP level during contraction, the level
can be maintained for a short time by
synthesis of ATP from creatine
phosphate and ADP.
• In a nonenzymatic reaction, small
amounts of creatine and creatine
phosphate cyclizes constantly to form
creatinine which can no longer be
phosphorylated and is therefore
excreted with the urine.
• Creatine does not derive from the muscles
themselves, but is synthesized in two steps in
the kidneys and liver.

• Initially, the guanidino group of arginine is

transferred to glycine in the kidneys, yielding
guanidino acetate.

• In the liver, N-methylation of guanidino acetate

leads to the formation of creatine from this.
• The coenzyme in this reaction is S-adenosyl
methionine (SAM).
SYNTHESIS OF CREATINE AND CREATININE
• MUSCLE METABOLISM 11
• A. cori and alanine cycle
• White muscle fibers mainly obtain ATP
from anaerobic glycolysis-i.e, they
convert glucose into lactate.
• The lactate arising in muscle and, in
smaller quantities, its precursor
pyruvate are released into the blood
and transported to the liver, where
lactate and pyruvate are resynthesize
into glucose again via
gluconeogenesis, with ATP being
consumed in the process.
• The glucose newly formed by the
liver returns via the blood to the
muscles, where it can be used as an
energy source again.
• This circulation system is called the
cori cycle.
• There is also a very similar cycle for
erythrocytes, which do not have
mitochondria and therefore produce
ATP by anaerobic glycolysis.
• The muscle themselves are not capable of
gluconeogenesis nor would this be useful, as
gluconeogenesis requires much more ATP than is
supplied by glycolysis.
• As 02 deficiencies do not arise in the liver even
during intensive muscle work, there is always
sufficient energy there, available for
gluconeogenesis.
• There is also a corresponding cycle for the amino
acid alanine.
• The alanine cycle in the liver not only provide
alanine as a precursor for gluconeogenesis, but
also transports the amino nitrogen arising in
muscles during protein degradation to the liver.

• In the liver, it is incorporated into urea for

excretion.
• Most of the amino acids that arise in
muscle during proteolysis are converted
into glutamate and -keto acids by
transamination.
• Again by transamination, glutamate and
pyruvate give rise to -ketoglutarate and
alanine, (which after glutamate/glutamine),
is the second important form of transport
for amino nitrogen in the blood.
• In the liver, alanine and ketoglutarate are
resynthesized into pyruvate and glutamate.
• Glutamate supplies ammonia to the urea
cycle, while pyruvate is available for
gluconeogenesis.
• B. protein and amino acid
metabolism
• The skeletal muscle is the most
important site for degradation of the
branched-chain amino acids (val, leu,
lle), but other amino acids are also
broken down in the muscles.
• Alanine and glutamate are
resynthesiszed from the components
and released into the blood.
• They transport the nitrogen that arises
during amino acid breakdown to the liver
(alanine cycle) and to the kidneys (as
glutamine).
• During periods of hunger, muscle proteins
serve as an energy reserve for the body.
• They are broken down into amino acids,
which are transported to the liver.
• In the liver, carbon skeletons of the amino
acids are converted into intermediates in the
tricarboxylic acid cycle (or into acetoacetyl-
CoA).
• These amphibolic metabolites are then
available for energy metabolism and for
gluconeogenesis.
• After prolonged starvation, the brain switches
to using ketone bodies (from acetyl-CoA
produced from fatty acids β-oxidation) in
order to save muscle protein.
• The synthesis and degradation of muscle
proteins are regulated by hormones.
• Cortisol leads to muscle degradation, while
testosterone stimulates protein formation.
• Synthetic anabolics with a testosterone-like
effect have repeatedly been used for doping
purposes or for intensive muscle-building.
• Note-In general for energy provision, red
muscles use fatty acids; ketone bodies also
serve as fuel especially for the heart muscles;
• white muscles use mainly glucose and in
resting muscle, fatty acids are the major
fuel.
• OXYGEN DEBT
• In actively contracting skeletal muscle,
the rate of glycolysis far exceeds that
of TCA cycle.
• Much of the pyrurate formed under
these conditions is reduced to lactate,
which flows to the liver, where it is
converted into glucose (via
gluconeogenesis).
• These interchanges, known as the cori
cycle shifts the metabolic burden of the
muscle to the liver.
• After a period of maximal muscular
exertion, such as a sprint, during which
lactate appears in the blood in large
amount, an animal will continue to
breath, in excess of the normal resting
state and consumes considerable extra
oxygen.
• The extra oxygen so consumed
during the recovery period is called the
“oxygen debt” and correspond to the
oxidation of some or all the excess
lactate formed during maximal
muscular contraction.
• Tetany and Rigor Mortis
• Tetany, a condition of hyper-contracted muscle that
sometimes follows a prolonged period of repetitive,
summed muscle stimulation, is caused by the depletion of
ATP and other high-energy phosphates that help maintain
normal ATP levels.
• The latter include other nucleoside triphosphates (NTPs),
creatine phosphate (CP), and ADP, as illustrated in the
three equations below.
• The three reactions are carried out by nucleoside
diphosphokinase, creatine kinase and adenylate kinase,
respectively.
• NTP + ADP → NDP + ATP
• CP + ADP → Creatine +ATP
• CONSEQUENCES OF TETANY
• Since tetanic stimulation raises sarcoplasmic calcium and
depletes ATP, the end result is a highly contracted muscle with
calcium bound to TnC and no ATP available to re-sequester
calcium into the cisternae of the SR, nor to break actomyosin
cross-bridges.
• Under these conditions, mitochondria will preferentially pump
calcium into the mitochondrial matrix, ultimately removing
calcium bound to TnC, obscuring myosin binding sites on thin
filaments, and, allowing the muscle to assume a flaccid state.
• However, the absence of ATP results in myosin remaining in its
low-energy conformational state, with the result that new
cycles of muscle stimulation will result in only limited ability of
the muscle to generate contractile activity.
• Muscles in this physiological state are said to be fatigued.
• RIGOR MORTIS
• In death, all reactions tend towards equilibrium.
• Among the first of these processes is that of ion equilibration
across all compartments of the body as ion pumps loose their
energy supplies.
• In the case of muscle, this results in cisternal and extracellular
calcium leaking into the sarcoplasm, raising calcium
concentrations to high levels.
• The calcium induces conformational changes in the troponin-
tropomyosin complex, exposing myosin binding sites on thin
filaments.
• The resulting uncontrolled contractile activity hastens the
total exhaustion of ATP supplies and ends with all or nearly all
myosin molecules in cross-linked actomyosin complexes.
• The rigid state of muscles that develops shortly after death is
due to this highly cross-linked state of thin and thick filaments