Journal of Muscle Research and Cell Motility 24: 593–596, 2003.
593
Ó 2003 Kluwer Academic Publishers. Printed in the Netherlands.
What is the role of tropomyosin in the regulation of muscle contraction?
The detailed nature of the interaction between actin and
myosin upon which striated muscle contraction depends
is not yet well defined nor is the precise mechanism by
which this interaction is regulated by the troponin
complex. Although molecular details of the calcium
switch are now available from the high resolution
structure of troponin C, we do not have similar
information about the other two components of the
complex, troponin I and T. Despite this uncertainty
tropomyosin has been considered for over 30 years to
have a well defined role in the regulation of contraction.
It is of interest to reflect why this is so for the evidence is
still circumstantial rather than direct. At the time the
steric hypothesis was proposed (Haselgrove, 1972;
Huxley, 1972; Parry and Squire, 1973), the X-ray
crystallographers and electron microscopists were mak-
ing the major contribution to the study of the structure
and function of the myofibril. When changes in the X-
ray diffraction pattern of the thin filament of intact
skeletal muscle during contraction were shown to be due
to movement of tropomyosin in the actin groove it was
logical to suggest that the former protein was involved
in the regulatory process. This hypothesis postulating
that tropomyosin ‘blocked’ the interaction of actin and
myosin in resting muscle but not in the stimulated state,
was an elegant and plausible explanation of the obser-
vations. A special role for tropomyosin in the contractile
system seemed likely for at that time contractile tissue
such as muscle and platelets appeared to be a unique
source of the protein. We now know that tropomyosin is
widely distributed in vertebrate tissue invariably associ-
ated with mature actin filaments. Muscle contains a
relatively large amount of the protein because the actin
thin filament system is a major component of the
cytoplasm. It is questionable, had we known in 1972 as
much about tropomyosin and its distribution as we
know today, whether the steric hypothesis, giving it a
unique role in muscle, would have been so widely
accepted.
It is beyond dispute that the tropomyosin molecule
moves on contraction in striated muscle but as yet the
mechanism responsible is not understood. In the models
incorporating a ‘blocking’ role for tropomyosin it is
usually suggested the conformational changes known to
arise when troponin C binds calcium are transmitted via
the troponin complex (Pearlstone and Smillie, 1983;
Phillips et al., 1986, Heeley et al., 1987; Kobayashi
et al., 2000). These involve troponin T, which binds to
tropomyosin, with result that the latter protein changes
its position in the groove of the actin filament to a ‘non-
blocking’ position. Tropomyosin is a rather flexible
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molecule and it is difficult to understand how confor-
mational changes occurring in the troponin C, close to a
point where the tropomyosin interacts with an actin
monomer, result in the whole tropomyosin molecule
moving in an identical manner with respect to the actin
surface extending over seven actin monomers.
Tropomyosin possesses two properties that are pre-
sumed to be of significance for its role in muscle and for
this reason are usually incorporated in models of
striated muscle regulation. (1) Its ability to interact with
actin. There are some unusual features in this interaction
and the factors which determine it are complex. Tropo-
myosin can inhibit, activate or have no effect on the
MgATPase of acto-heavy meromyosin, depending on
the ionic strength, the isoforms present, temperature, the
ratio of actin to myosin heads and the presence of other
myofibrillar proteins (see Perry, 2001, for review). (2) Its
ability to endow cooperative properties on the F-actin
filament. These are clearly shown by its effects on the
behaviour of myosin MgATPase (Bremel et al., 1972)
and on the inhibitory properties of troponin I (Perry
et al., 1972).
Interaction with actin is vital for a simple ‘blocking’
mechanism from which cooperativity may or may not
arise depending on the model proposed. It is now clear
from several lines of evidence that in the ‘blocked’ or
resting state some interaction between actin and myosin
can take place, but not that which leads to the high rate
of MgATPase associated with contraction (Chalovich
et al., 1981; Stein et al., 1981; Resetar et al., 2002; see
Chalovich, 2002). Electron microscope reconstruction
studies of the position of tropomyosin on the actin
filament in the ‘blocked’ state (Vibert et al., 1997; Xu
et al., 1999; Craig and Lehman, 2001) indicate that part
of the putative myosin binding site(s) is exposed. In
consequence the original ‘blocking’ mechanism has been
modified to accommodate these observations.
The conclusion that tropomyosin ‘blocks’ the inter-
action of actin and myosin because in the resting state
the position of tropomyosin on actin is close to those
regions presumed to interact with myosin may not be
justified. The interaction between actin and myosin is
clearly complex with a number of contact points that
may be involved in a coordinated manner and which are
not well defined. Most workers agree that on contrac-
tion troponin moves with the tropomyosin but there is
still some uncertainty about its contribution to the X-
ray data. It has, however, been distinguished from the
latter in electron microscope reconstruction studies
(Lehman et al., 2001). Nevertheless these studies have
so far failed to prove it is the tropomyosin rather than
the troponin I that is binding to actin to prevent
interaction in the resting state. The troponin complex
594
extends alongside the tropomyosin occupying the C
terminal half of the molecule with troponin I and C
located in a more central position with respect to the
tropomyosin molecule (Perry, 1998). The experimental
evidence we have from in vitro studies suggests that in
the absence of calcium i.e. in the ‘blocked’ state the
troponin I is bound to actin. As the calcium concentra-
tion rises the affinity of troponin C for troponin I
increases markedly. The inhibitory peptide site of
troponin I detaches from actin, troponin I binds to
troponin C and the actin monomers return to their
ground state conformation in which they can activate
the myosin MgATPase.
Recent NMR and fluorescence studies (Patchell et al.,
2002; Patchell, Perry and Levine, unpublished), indicate
that peptides corresponding to sites on human cardiac
ß-myosin suggested from models as possible interaction
sites, e.g. residues 398–414 and 622–646, bind to actin.
In the presence of troponin I, or the inhibitory peptide
derived from it, these myosin peptides dissociate from
actin. The evidence indicates that the myosin peptides
and the inhibitory peptide do not occupy identical sites
on actin and implies that interaction with troponin I
induces a conformational change in actin so that it is
unable to interact with myosin in a manner that
activates the MgATPase. Tropomyosin did not block
the binding of the myosin peptides; indeed it strength-
ened their affinity for actin. It also strengthened the
affinity of the inhibitory peptide for actin and in this way
increased its ability to impose a conformational change
on the actin.
There was no evidence that tropomyosin alone
inhibited the binding of any of the myosin peptides
tested. These investigations must be extended to myosin
subfragment 1 in the nucleotide bound states for further
definition of the system. Nevertheless extrapolation
from the results obtained with peptide segments suggest
that in the ‘blocked’ state interaction with these regions
of the myosin molecule is prevented by a conformational
change imposed on actin by troponin I, not by the
‘blocking’ action of tropomyosin.
The functional unit of the thin filament consists of
seven actin monomers, one molecule of tropomyosin
TNI
TNI
Fig. 1. Diagrammatic representation indicating how conformational changes induced in actin by troponin I (TNI) could be transmitted to
neighbouring monomers in the thin filament (thin arrows). For the purpose of representation the actin duplex is uncoiled. Position of
tropomyosin indicated by a thick black line in resting muscle and a hatched line in contracting muscle.
and troponin composed of one molecule each of
troponin I, C and T. If tropomyosin really ‘blocks’ a
site on actin essential for activation it is easy to visualise
it lying alongside the actin filament close to the
appropriate sites on each of seven actin monomers.
The whole functional unit would be in effect under the
control of one tropomyosin molecule, as the steric
hypothesis implies. On the other hand, if as the more
recent direct evidence suggests, troponin I interacts with
actin to produce the conformational change that pre-
vents interaction with myosin, only one actin monomer
(possibly two) will be directly involved. It is suggested
that in the presence of tropomyosin the conformational
changes imposed by troponin I on one actin monomer
are transmitted to monomers on either side in the
filament. If these effects were transmitted to three
adjacent actin monomers on each side of the troponin
I interaction site the whole functional unit would be
controlled by changes occurring on the actin monomer
that binds troponin I (Figure 1). In view of the evidence
that possibly two actin monomers are involved at the
site of interaction with myosin, one troponin I molecule
may directly effect two actin monomers. If this were the
case the transmission of conformational changes be-
tween monomers would be aided.
The structure of the double helical F-actin filament is
stabilised by many interactions between monomers in
the single filaments and between monomers in different
filaments of the duplex. It is likely that conformational
changes imposed on single monomers will induce
changes in adjacent monomers to maintain the double
filament structure. The fact that troponin I molecules
are located opposite each other on the adjacent actin
filaments of the duplex (Figure 1) probably strengthens
the effect of troponin I and the transmission of the
conformational change along the actin duplex. The role
of tropomyosin would be to stabilise the filament
structure and thus facilitate the transmission of confor-
mational changes along the filament. There are many
reports of the stabilising and stiffening effect of tropo-
myosin on the actin filament (Kawamura and Maru-
yama, 1970; Fujime and Ishiwata, 1971; Kojima et al.,
1994; Goldmann, 2000; Kuo-Kang et al., 2000). It is
TNI TNI
TNI TNI

clear that on binding, tropomyosin imposes some effect
on the structure of actin for in its presence the affinities
of troponin I inhibitory peptides, myosin peptides
(Patchell et al., 2002), troponin (Wegner and Walsh,
1981) and myosin (Bremel et al., 1972; Eaton, 1976) are
increased.
If the conformational changes in actin are transmitted
along the filament, changes will occur in the distribution
of residues on its surface. Some of these residues will be
involved in the binding to tropomyosin and the fibrous
molecule will have to adjust its position along its whole
length on the F-actin surface for energetic consider-
ations. The interaction between actin and tropomyosin
is rather unusual, certainly different from that between
actin and troponin I where specific sites are involved
(Grabarek and Gergley, 1987; Levine et al., 1988).
Tropomyosin interacts with different actin monomers
via each of the quasi-equivalent regions which although
similar are not identical in sequence. The distance
between the carbon atoms of actin and tropomyosin in
the model of the complex proposed by Lorenz et al.
(1995) is about 10 Å which implies that the interaction is
mainly ionic as the effects of magnesium and potassium
chloride on complex formation suggest. With tropomyo-
sin floating as it were in the actin groove its position is
determined largely by ionic interactions. Conformation-
al changes on the surface of the actin monomer
probably involving both hydrophilic and hydrophobic
side chains would lead to a change in the position of the
tropomyosin molecule.
The role of tropomyosin in inducing cooperativity
into the actin filaments is illustrated by the use of in vitro
systems of actomyosin and troponin I. Addition of
tropomyosin converts the effect of troponin I to produce
maximum inhibition of the MgATPase from a molar
ratio of troponin I to actin of approximately 1:1 to
approximately 1:7 (Perry et al., 1972; Lehrer et al.,
1997; Geeves et al., 2000). Thus in the presence of
tropomyosin and troponin I, but in the absence of the
other components of the troponin complex, no actin
monomer in the functional unit is able to fully activate
the myosin MgATPase. The potentiation of the inhi-
bitory effect of the intact molecule of troponin I is also
obtained with 21 and 12 residue peptides (Syska et al.,
1976; Talbot and Hodges, 1981). It is difficult to
envisage a mechanism that enables such small peptide
regions of troponin I to displace tropomyosin into an
inhibitory position along the whole of its length as a
‘blocking’ role would demand. A more plausible expla-
nation would be that tropomyosin facilitates the trans-
mission of the effect of the conformational change
imposed by troponin I peptide on actin to all the
monomers in the functional unit.
Until the atomic details of the actomyosin interface
and nature of the interaction of the troponin complex
with it have been defined at high resolution it is not
possible to be certain about the role of tropomyosin in
the regulatory process. In this discussion attention has
been focussed on striated muscle where it is becoming
595
clear that conformational changes imposed on actin by
the unique inhibitory protein of striated muscle, tro-
ponin I, are of particular significance for regulation.
Movement of tropomyosin also occurs in smooth
muscle although its location in the actin groove is
different to that in striated muscle (Lehman et al., 1996).
Myosin and particularly actin are strongly conserved
proteins and despite the differences in the regulatory
system in smooth muscle, the role of tropomyosin in
relation to actin would be expected to be similar to that
in striated muscle. The difficulties in ascribing a ‘block-
ing’ role for tropomyosin in smooth muscle regulation
are discussed elsewhere (Marston et al., 1998).
If the role of tropomyosin is to facilitate the trans-
mission of conformational changes between actin mono-
mers the difficulties in explaining its role in all types of
muscle disappear. Such a general role makes good
functional sense for actin-binding proteins with different
roles are widely distributed as are actin filaments. Actin
filaments are built up of a large number of subunits
usually much more abundant than the functional
proteins that bind to them. It is biologically efficient
for the conformational effects induced by the binding
proteins on the actin filament to be transmitted to the
actin monomers with which they are not in contact.
The tropomyosin molecule polymerised as a continuous
filament in each of the two grooves of the actin duplex is
ideally located for this role. The proposed mechanism
of function implies that the role of tropomyosin is
not a special case unique to muscle but merely an
example of the role played in all actin filament systems.
It is suggested that striated muscle is regulated by a
specialised allosteric switch applied by troponin I on
actin and facilitated by the general properties of
tropomyosin.
I am grateful to Val Patchell, Barry Levine, Emil
Toescu and Joe Chalovich who have made helpful
comments on this article.
S.V. Perry
Department of Physiology
Division of Medical Sciences
Medical School
University of Birmingham
Edgbaston
Birmingham B15 2TT
UK
References
Bremel RD, Murray JM and Weber A (1972) Manifestations of
cooperative behaviour in the regulated actin filament during actin-
activated ATP hydrolysis in the presence of calcium. Cold Spring
Harbor Symp Quant Biol 37: 267–265.
Chalovich JM (2002) Regulation of striated muscle contraction: a
discussion. J Muscle Res Cell Motil 23: 353–361.
Chalovich JM, Chock PB and Eisenberg E (1981) Mechanism of action
of troponin–tropomyosin inhibition of actomyosin ATPase activ-
ity without inhibition of myosin binding to actin. J Biol Chem 256:
575–578.
596
Craig R and Lehman W (2001) Cross bridge and tropomyosin
positions observed in native interacting thick and thin filaments. J
Mol Biol 311: 1027–1036.
Eaton BL (1976) Tropomyosin binding to F-actin induced by myosin
heads. Science 192: 1337–1339.
Fujime S and Ishiwata S (1971) Dynamic study of F-actin by
quasielectric scattering of laser light. J Mol Biol 62: 251–265.
Geeves MA, Chai M and Lehrer SS (2000) Inhibition of actin–myosin
ATPase activity by troponin I and IC: relationship to the thin
filament states of muscle. Biochemistry 39: 9345–9350.
Goldmann WH (2000) Binding of tropomyosin–troponin to actin
increases filament bending stiffness. Biochem Biophys Res Commun
276: 1225–1228.
Grabarek Z and Gergely J (1987) Troponin–I binds to the N-terminal
12-residue segment of actin. Biophys J 51: A331.
Haselgrove JC (1972) X-ray evidence for a conformational change in
the actin containing filaments of vertebrate striated muscle. Cold
Spring Harbor Symp Quant Biol 37: 341–352.
Heeley DH, Golosinska K and Smillie LB (1987) The effects of
troponin T fragments T1 and T2 on the binding of non-
polymerisable tropomyosin to F-actin in the presence and absence
of troponin I and troponin C. J Biol Chem 262: 9971–9978.
Huxley HE (1972) Structural changes in the actin- and myosin-
containing filaments during contraction. Cold Spring Harbor Symp
Quant Biol 37: 361–376.
Kawamura M and Maruyama K (1970) Electron microscopic particle
length of F-actin in vitro. J Biochem 67: 437–457.
Kobayashi T, Kobayashi M, Gryczynski Z, Lakowitz R and Collins JH
(2000) Inhibitory region of troponin: Ca2+ dependent structural
and environmental changes in the troponin–tropomyosin complex
and in reconstituted thin filaments. Biochemistry 39: 86–91.
Kojima H, Ishijima A and Yanagida Y (1994) Direct measurement of
stiffness of single actin filaments with and without tropomyosin by
in vitro nanomanipulation. Proc Natl Acad Sci USA 91: 12962–
129.
Kuo-Kang W, Kang B and Rubenstein PA (2000) Tropomyosin-
dependent filament formation by a polymerization-defective mu-
tant yeast actin (V266G, L267G). J Biol Chem 275: 40594–40600.
Lehman W, Vibert P, Craig R and Barany M (1996) Actin and the
structure of smooth muscle thin filaments. In: M. Barany (ed.)
Biochemistry of Smooth Muscle Contraction. (pp. 47–60) Academic
Press, New York.
Lehman W, Rosol M, Tobacman LS and Craig R (2001) Troponin
organisation on relaxed and activated thin filaments revealed by
electron microscopy and three dimension reconstruction. J Mol
Biol 307: 739–744.
Lehrer SS, Chai M and Geeves MA (1997) Effect of troponin I (TNI)
on actin S1 ATPase and S1 binding in the absence and presence of
rabbit skeletal tropomyosin. Biophys J 72: MP 198.
Levine BA, Moir AJG and Perry SV (1988) The interaction of
troponin-I with the N-terminal region of actin. Eur J Biochem 172:
389–397.
Lorenz M, Poole KJV, Popp D, Rosenbaum G and Holmes KC (1995)
An atomic model of the unregulated thin filament obtained by X-
ray fiber diffraction on oriented actin–tropomyosin gels. J Mol Biol
246: 108–119.
Marston SB, Burton D, Copeland O, Fraser A, Gao Y, Hodgkinson P,
Huber PA, Levine B, El-Mezgueldi M and Notoriani G (1998)
Structural interactions between actin, tropomyosin, caldesmon and
calcium binding protein and the regulation of smooth muscle thin
filaments. Acta Physiol Scand 164: 401–414.
Parry DAD and Squire JM (1973) Structural role of tropomyosin in
muscle regulation: analysis of the X-ray diffraction patterns for
relaxed and contracting muscles. J Mol Biol 75: 35–55.
Patchell VB, Gallon CE, Hodgkin MA, Fattoum A, Perry SV and
Levine BA (2002) The inhibitory region of troponin I alters the
ability of F-actin to interact with different segments of myosin. Eur
J Biochem 269: 5088–5100.
Pearlstone JR and Smillie LS (1983) Effects of troponin I plus C on the
binding of troponin T and its fragments to a-tropomyosin. J Biol
Chem 258: 2534–2542.
Perry SV (1998) Troponin T: genetics, properties and function. J
Muscle Res Cell Motil 19: 575–602.
Perry SV (2001) Vertebrate tropomyosin: distribution, properties and
function. J Muscle Res Cell Motil 22: 5–49.
Perry SV, Cole HA, Head JF and Wilson FJ (1972) Localisation
and mode of action of the inhibitory component of the tro-
ponin complex. Cold Spring Harbor Symp Quant Biol 37: 251–262.
Phillips GN Jr., Filler JP and Cohen C (1986) Tropomyosin crystal
structure and muscle regulation. J Mol Biol 192: 111–131.
Resetar AM, Stephens JM and Chalovich JM (2002) Troponin–
tropomyosin: an allosteric switch or steric blocker. Biophys J 83:
1039–1049.
Stein LA, Chock FB and Eisenberg E (1981) Mechanism of the
actomyosin ATPase: effect of actin on the ATP hydrolysis step.
Proc Natl Acad Sci USA 78: 1346–1350.
Syska H, Wilkinson JM and Perry SV (1976) The relationship between
biological activity and primary structure of troponin I of white
skeletal muscle of the rabbit. Biochem J 153: 375–387.
Talbot JA and Hodges RS (1981) Synthetic studies on the inhibitory
region of rabbit skeletal troponin I. J Biol Chem 256: 2798–2802.
Vibert P, Craig R and Lehman W (1997) Steric-model for activation
of muscle thin filaments. J Mol Biol 266: 8–14.
Wegner A and Walsh TP (1981) Interaction of tropomyosin–troponin
with actin filaments. Biochemistry 20: 5633–5642.
Xu C, Craig R, Tobacman L, Horowitz R and Lehman W (1999)
Tropomyosin positions in regulated thin filaments revealed by
cryoelectron microscopy. Biophys J 77: 985–992.