TOPIC 2

Natural Muscles

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 CHAPTER 2
Natural Muscle as a Biological System
Gerald H. Pollack, Felix A. Blyakhman, Frederick B. Reitz,
Olga V. Yakovenko, and Dwayne L. Dunaway
University of Washington

2.1 Conceptual Background / 53

2.2 Structural Considerations / 56
2.3 Does Contraction Involve a Phase Transition? / 59
2.4 Molecular Basis of the Phase Transition / 62
2.4.1 Connecting Filaments / 62
2.4.2 Thick Filaments / 64
2.4.3 Thin Filaments / 65
2.5 Lessons from the Natural Muscle System That May Be
Useful for the Design of Polymer Actuators / 68
2.6 References / 70

Muscle is the natural contractile system that artificial systems attempt to emulate.
For proper emulation it is evidently necessary to understand the basic underlying
mechanism. In the material that follows, we consider the evidence that muscle is
a polymer gel, and that the gel’s polymeric filaments contract by a polymer-gel
phase-transition. This is an unorthodox conclusion, and we therefore begin by
considering the relevant background.

2.1 Conceptual Background

Until the mid-1950s muscle contraction was held to occur by a protein-folding
mechanism [for review, cf. A. F. Huxley, 1957]. The folding mechanism is
similar enough to the one that will be suggested here that it may be considered a
progenitor.
With the discovery of interdigitating thick and thin filaments in the mid-
1950s, it was tempting to discard the notion of folding, and suppose instead that
contraction arose out of pure filament sliding. This supposition led H. E. Huxley
and Sir Andrew Huxley to examine independently whether the interdigitating
filaments (Fig. 1) remained at constant length during contraction. Their optical
microscopic studies of muscle-striation patterns were published back-to-back in
Nature [Huxley and Hanson, 1954; Huxley and Niedergerke, 1954]. Together

53

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 54 Chapter 2

with more detailed follow up [Huxley and Niedergerke, 1958], these studies
became the field’s pivotal works.
These studies examined relaxed and activated specimens alike. Relaxed
specimens were manually stretched and released. During these maneuvers, the
width of the A-band remained absolutely invariant. Since A-band width
corresponds to the length of the thick filament (Fig. 1), the result implied that
thick filaments did not change length. This strengthened the emerging notion of
filament sliding.

Figure 1: Molecular structure of the sarcomere. Cartoon of a microscopic view

(above) corresponds to set of interdigitating filaments. Thick filaments
correspond to dark A-band. Corresponding molecular structure (below) shows
connecting filaments in series with thick filaments. Thin filaments lie in parallel
with thick and connecting filaments.

Less conclusive, however, were the results obtained when muscle length
changed actively, for in this case sarcomere shortening was sometimes
accompanied by A-band narrowing. The narrowing was dismissed as
quantitatively inconsequential in the Huxley–Hanson study, and in the Huxley–
Niedergerke study it was relegated to the microscope’s limited resolution. With
these dismissals, all results could remain interpretable within the framework of
constant-length filaments.
These back-to-back papers by the Huxleys dispatched the protein-folding
paradigm to the heap of forgotten notions. The constant filament-length paradigm
took hold, and has held remarkably firm ever since—notwithstanding subsequent

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 Natural Muscle as a Biological System 55

findings of thick filament or A-band shortening in more than 30 reports [for

review, see Pollack, 1983; Pollack, 1990].
With the emerging notion of sliding filaments, the central issue became the
nature of the driving force. Impressive numbers of researchers sought out a
mechanism, and soon settled on the one schematized in Fig. 2. In this
mechanism, the cross-bridges attach to the thin filament, swing, and then detach
in the presence of ATP, readying themselves for the next cycle. Bridge swinging
drives the thin filament toward the center of the sarcomere, thereby shortening
the sarcomere and the muscle. The mechanism explains many known features of
contraction and has therefore become broadly accepted [for review, cf. Spudich,
1994; H. E. Huxley, 1996; Block, 1996; Howard, 1997; Cooke, 1997].

Figure 2: Cross-bridge model of muscle contraction. In the relaxed state (top)

bridges projecting from thick filament do not interact appreciably with the thin
filament. In the activated state (bottom), bridges attach and rotate, thereby
driving the thin filament rightward, toward the center of the sarcomere. Z-line is at
left.

Apart from the concern of building on the uncertain foundation of constant-

length filaments, a serious concern is the absence of evidence for cross-bridge
swinging. To test for bridge-angle changes, numerous molecular scale
approaches have been applied [for review, see Thomas, 1987]. Electron-spin
resonance, x-ray diffraction, and fluorescence polarization have produced largely
negative results, as has high-resolution electron microscopy [Katayama, 1998].
Of all these results, the most supportive has been an angle change of 3 deg
measured on a cross-bridge (myosin-light chain) appendage [Irving et al., 1995].
This 3 deg change is still far short of the anticipated 45 deg.

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 56 Chapter 2

Absence of swinging is only one of several areas of concern. Concerns run

the gamut from instability [Iwazumi, 1970] to mechanics [Pollack, 1983],
structure [Schutt and Lindberg, 1993, 1998], and chemistry [Oplatka, 1996,
1997]. Many of these concerns are detailed in my 1990 book [Pollack, 1990]. But
the full flavor is best gleaned by reading the original reviews, particularly those
by Schutt and Lindberg and by Oplatka, whose spicy, no-holds-barred bluntness
entertains as it educates. I also commend an insightful work by the late Graham
Hoyle [1983], in which some understanding of the field’s continued focus on
seemingly unproductive paradigms is offered in a chapter labeled “Why Do
Muscle Scientists ‘Lose’ Knowledge?”
In the material that follows, we pick up where the pioneers of a half-century
ago who championed the protein folding mechanism, including Nobelist Albert
Szent-Györgyi, left off. We first deal with several relevant structural
considerations, and then with possible mechanisms, keeping a sharp eye toward
relevance for artificial muscles.

2.2 Structural Considerations

The functional unit of contraction is the myofibril, which comprises several
hundred protein filaments (Fig. 3). Many parallel myofibrils make up the muscle
fiber, or cell; and many muscle fibers make up the muscle. The arrangement is
hierarchical.
The myofibrillar sarcomere comprises three types of polymeric filament, two
of which were illustrated in Fig. 2. The central one is the thick filament, which is
built of multiple repeats of the protein myosin. The thick filament is flanked at
either end by a connecting filament, which in turn connects to the Z-line. In
vertebrate muscle the connecting filament is built largely of the protein titin—a
polymer made up principally of tandem repeats of discrete immunoglobulinlike
domains. The thin filament is also polymeric. Along with various regulatory
proteins, it consists mainly of multiple repeats of the protein actin. Thus, all three
of the sarcomere’s longitudinal structures are biological polymers.
The myofibril is striking in the almost crystalline precision with which
filaments align with one another in parallel, conferring sharpness on the A-I
boundaries. Such alignment is realized through extensive cross-linking
throughout the sarcomere, both in the A-band and in the I-band.
I-band cross-links are illustrated in Fig. 4. The top panel is a freeze-fracture
image, which reveals the cross-links in lifelike form. The bottom panel is a
conventional thin section. Cross-linking is a general rule among diverse actin
filaments, and therefore it is no surprise that it occurs in the sarcomere.

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 Natural Muscle as a Biological System 57

Figure 3: Structure of an intact myofibril. The A-band contains a parallel array of

thick filaments. Connecting filaments lie in the I-band. Thin filaments originate at
the Z-line and partially overlap thick filaments in the A-band.

Figure 4: Top: Freeze fracture image of I-band filaments. Vertically oriented

mesh at left is the Z-line. Arrow indicates interconnection. Bottom: Ultrathin
section showing regular repeat of I-band interconnections. View from shallow
angle, below. (Reprinted from Baatsen et al. 1988. Copyright 1988, with
permission from Elsevier.)

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 58 Chapter 2

Figure 5: Top: Thin section of honeybee flight muscle in which thin filaments had
been withdrawn to increase clarity of thick filament zone. Arrows denote
interconnections. Bottom: cross-section through A-band of moderately stretched
sarcomere that grazed tips of thin filaments, the latter seen as occasional small
dots. Interconnections between thick filaments are evident.

A-band interconnections are shown in Fig. 5. The top panel shows a

specimen that had been stretched by about 30% to withdraw thin filaments from
the lattice in order to reduce visual congestion. Stretch distorts the lattice
somewhat, but cross-links between thick filaments nevertheless remain clear.
The thick-to-thick interconnections are almost certainly built of myosin, for
myosin is the only A-band protein in sufficient quantity to account for structures
so abundant. Links of appropriate length can be created if cross-bridges
extending from adjacent filaments bind to one another at their tips [Pollack,
1990]. The cross-links visible in these micrographs are not widely recognized in
the muscle field, where free, swingable cross-bridges are required for the theory.

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 Natural Muscle as a Biological System 59

On the other hand, these essentially static elements may explain why attempts to
detect myosin-head tilting have failed; rungs cannot tilt.
Cross-links align the filaments and thereby confer regularity on the lattice;
that is why the A–I junctions are so distinct. They also maintain lattice integrity
by limiting swelling. Highly charged polymers such as those in the sarcomere
can imbibe water up to thousands of times their volume [Osada and Gong, 1993],
and may ultimately dissolve if not cross-linked. The muscle-filament lattice,
then, is essentially a highly cross-linked, water-filled polymer gel.

2.3 Does Contraction Involve a Phase Transition?

This gel is evidently designed to contract, and certain features of its behavior
imply a phase transition. Triggers of contraction, for one, are the same as those
for transitions in ordinary polymer-gels [Hoffman, 1991]. In contractile protein
bundles of demembranated cells, for example, contraction can be initiated by
increased salt, pH change, temperature jump—even electrical current will trigger
contraction of an isolated myofibril [Garamvolgyi, 1959]. And like polymer-gel
transitions, triggering is critical, or “razor-edge” (see below). Nothing happens
until a threshold is crossed, whereupon contractile action is massive.
Ironically, such critical behavior had been evident in model studies carried
out more than a half-century ago, and that is perhaps the reason why contraction
was presumed to involve something akin to a phase transition. Vanguard
experiments of the era focused on suspensions of actin and myosin. Such
suspensions could form a gel, the condensation of which was considered a
working model of muscle contraction. The gel remained stable until ambient
conditions were edged just past a critical threshold; then it contracted massively.
As it contracted, the matrix folded and water was released—much the same as it
is released in the polymer-gel (Fig. 6).

Figure 6: The actomyosin gel undergoes massive volume change in response to

a slight increase of ATP concentration. (Reprinted from Szent-Györgyi, 1951.
Copyright 1951 with permission from Elsevier.)

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 60 Chapter 2

Like the polymer gel, the cross-linked polymer network of the sarcomere is
invested with solvent—an aqueous salt solution. The solvent remains trapped
within the network; it does not leak out. This can be seen in specimens whose
membranes have been removed (skinned fibers): such specimens do not easily
lose solvent. Again, this phenomenon is similar to that of the gel, the solvent
presumably held by strong hydrophilic forces that retain the water within the
sarcomere’s polymeric framework [Pollack, 2001]. Thus, muscle exhibits the
essential features of polymer gels—except that its polymers are well organized
into a regular framework and not randomly dispersed.
If muscle contraction were to involve a phase transition, the resulting
dimensional change would be axial because polymers are naturally aligned along
the axis of the muscle; hence, polymer shortening would produce shortening
along the axis of the muscle. On the other hand, polymeric fragments of actin and
myosin can be used to construct random or semi-random gels, in which case
contraction should be isotropic, as in the gel of Fig. 6, above. The behavior
illustrated in Fig. 6 is rather similar to the behavior of polyacrylamide gels
exposed to varying ratios of organic/inorganic solvents [Tanaka, 1981]. Not only
is contraction fairly isotropic, but it also has a critical nature: no change at all
until the stimulus reaches a threshold, and then massive condensation.
Critical behavior is also seen when muscle polymers are naturally oriented,
as they are in striated muscle. In Fig. 7 the concentration of calcium, the
physiological trigger, is progressively increased. Stripped of its membrane, the
contractile specimen is immersed in a physiological solution in which the level of
free calcium is progressively elevated. Once the concentration crosses threshold,
full tension is observed within a narrow concentration window. Similar behavior
is observed when calcium is replaced by other divalent cations such as barium or
strontium. Contraction is essentially all-or-none.
Critical behavior is also seen when an organic solvent surrounding the
specimen is progressively replaced by an aqueous solvent; see Fig. 8. Again, the
contraction is largely all-or-none, within a narrow window of organic/aqueous
solvent ratio. Such behavior is a classical signature of a polymer gel phase-
transition. Thus, critical “razor-edge” behavior is preserved in the naturally
oriented polymeric system just as it is in the random muscle gel.
The evidence above implies that muscle contracts in the same way as an
ordinary polymer gel contracts. How, then, might such behavior be manifested at
the molecular level? The evidence is most consistent with a mechanism in which
the polymers themselves can contract. In fact, evidence in the literature, as shown
below, implies that all three polymers can shorten.

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 Natural Muscle as a Biological System 61

Figure 7: Effect of increase of calcium concentration on relative isometric tension

development in single rabbit muscle cells [Pollack, 1990]. (Courtesy of Ebner &
Sons.)

Figure 8: Effect of solvent variation on contraction in rabbit muscle [Pollack,

1990]. (Courtesy of Ebner & Sons.)

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 62 Chapter 2

2.4 Molecular Basis of the Phase Transition

In the proposed paradigm, all three filaments shorten by a phase transition, and
this shortens the sarcomere. I will briefly sketch some of the evidence that leads
to this kind of mechanistic view, considering filaments one at a time. The
paradigm suggested here is outlined in Pollack [1990], with some newer elements
considered in a more recent review article [Pollack, 1996].

2.4.1 Connecting Filaments

Connecting filaments are springlike polymers that retract when stretched. When
unactivated muscles are forcibly stretched, the connecting filaments are strained.
If the muscle is then released, connecting filaments retract and their tension is
relieved. Connecting filaments are thereby responsible for the muscle’s so-called
resting tension. Resting tension ensures that the stretched sarcomere returns to its
natural length, and also keeps the thick filament centered in the sarcomere. All of
these considerations refer to muscles that are not stimulated to contract actively;
thus, the force range under consideration is relatively modest.
Dynamics of the connecting filament have been revealed in elegant
experiments on isolated molecules of titin [Rief et al., 1997; Tskhovrebova et al.,
1997]. In the Rief et al. study, ramp-length changes imposed on single titin
molecules elicited sawtoothlike tension changes. This implied a one-by-one
unfolding of the filament’s tandem immunoglobulinlike domains—each
unfolding event giving rise to one “tooth” in the sawtooth-tension waveform. The
length change occurs as each immunoglobulin “beta-barrel” domain gives way to
an extended random structure (Fig. 9). Many such unfoldings lengthen the
filament in steplike fashion.
In a conceptually similar experiment [Tskhovrebova et al., 1997], single titin
molecules were stretched and held. During the holding phase, the decline of
tension (stress-relaxation) occurred in stepwise fashion, not smoothly. Again, the
implication is a progression of discrete unfolding events, each one dropping the
tension by an increment.

Figure 9: Unfolding of immunoglobulin domain (left to right) results in discrete

length change. Unfolding of successive domains yields a large-scale length
change of the connecting filament.

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 Natural Muscle as a Biological System 63

Stepwise length changes are also observed under similar conditions in

sarcomeres [Yang et al., 1998]. These experiments interrogated the single
myofibril, the smallest functional unit that retains muscle’s natural structure (see
Fig. 3). When the unactivated myofibril is stretched or released by a linear ramp,
the sarcomere-length change is stepwise. An example is shown in Fig. 10.

Figure 10: Time course of sarcomere-length change during smooth trapezoidal

length change imposed on single isolated myofibril. Calibration bars: 20 nm; 1
sec.

Myofibril sarcomere steps reveal that discrete behavior persists at levels of

organization considerably higher than that of the single molecule, for the
myofibrillar sarcomere contains hundreds of filaments in parallel. In fact, discrete
behavior remains detectable all the way up to the cellular level [Granzier et al.,
1985], reinforcing the highly cooperative nature of the step.
The step’s character implies a phase transition. First, the onset and
termination are abrupt—distinct states appear to be involved. Second, the step is
not an isolated event: the fact that it occurs over many different molecules at the
same time implies a quasi-all-or-none behavior, which again is characteristic of a
phase transition. The simplest interpretation is that the phase-transition
progresses collectively along all parallel connecting filaments, shortening or
lengthening by one domain at a time.

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 64 Chapter 2

2.4.2 Thick Filaments

The thick filament is built of successive repeats of the myosin molecule. Myosin
contains a long alpha-helical rodlike tail, most of which lies in the thick filament
backbone (extensions of the angled lines in Fig. 2); it also contains two globular
heads, or cross-bridges, which radiate from the filament backbone at nominally
right angles. The rodlike tails overlap in a staggered, helical pattern. The filament
is mirror-symmetrical about its midpoint.
Although it is generally accepted that thick filaments do not change length
during contraction, the evidence is mixed. Classical studies imply a constancy of
filament length under most conditions. But some thirty-plus papers published
since those classical studies report substantial shortening during contraction [for
review, cf. Pollack, 1983]. These studies were conducted on vertebrate heart and
skeletal muscles as well as invertebrate muscles; they employed techniques
ranging from electron microscopy through various types of optical microscopy,
and they included isolated thick filaments extracted from the muscle and exposed
to physiological activating agents. The evidence for thick-filament shortening is
therefore appreciable.
A way in which thick filaments might shorten is if myosin rods were able to
slide past one another. The filament could then “telescope” to a shorter length.
The driving force for such telescopic action would lie in the filament itself,
presumably in the myosin rods that comprise the backbone. In this context it has
been shown in many experiments that the alpha-helical rod is able to shorten by a
helix-coil transition [for review, cf. Pollack, 1990]. The helix-coil transition is a
phase transition in which the molecular structure undergoes radical change. It is a
force-producing process: because the equilibrium length of the random coil is
near zero, the coil will always wants to shorten from its extended length. The
retractive force depends on the degree of extension, much like a spring. Thus, the
transition shortens the myosin molecule in much the same way as a wool sweater
is shortened by excess heat. Here, however, the transition is reversible.
A model illustrating how localized shortening in a myosin rod could generate
rod sliding is shown in Fig. 11. Adjacent molecules are held together by cohesive
forces derived from molecular surface charges that alternate semi-regularly along
the surface. The unstable zone of one molecule lying near the filament’s
midpoint undergoes a helix-coil transition. (The transitions would occur
symmetrically about the filament’s midpoint.) The transition results in local
shortening, which draws the remainder of the filament toward the mid-zone.
Then the same shortening event occurs in the next molecule along the filament,
and the next, etc. In such a way the filament shortens step by step, on either side
of the midpoint. And because molecules are cross-linked to respective molecules
on the next filament in parallel, the transition is cooperative over the muscle
cross section.

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 Natural Muscle as a Biological System 65

Figure 11: Mechanism by which local shortening in one myosin rod can propel
the adjacent rod to slide. Only two of the many molecules are shown.

This mechanism is in good agreement with structural evidence from x-ray

diffraction [Huxley and Brown, 1967; Yagi and Matsubara, 1984]. The x-ray
patterns show that during active sarcomere shortening the spacing between
molecules along the thick filament does not change; only the x-ray intensity
changes. This is precisely what is anticipated. The x-ray pattern is dominated by
contributions from those molecules along the thick filament that remain regularly
arrayed and have not yet shortened; others have shortened by variable amounts
and do not contribute significantly. As the number of transitioned molecules
increases, their intensity contribution therefore diminishes, explaining the
observed x-ray intensity diminution. The model is therefore in good agreement
with ultrastructural and x-ray diffraction evidence.
If thick filaments shorten during contraction, the likelihood, then, is that such
shortening occurs in steps, one molecule at a time in each half-filament. If the
thin filament is bound to the cross-bridges during these thick filament-shortening
steps, the sarcomere will likewise shorten in steps. In such a way the myosin
phase transition contributes to the shortening of the sarcomere.

2.4.3 Thin Filaments

Thin filaments may bring about stepwise length changes as well. Unlike the thick
and connecting filaments, whose shortening can directly shorten the sarcomere,

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 66 Chapter 2

thin filaments are differently situated (Fig. 1). Because of their arrangement, they
would need to facilitate contraction in a different way. One potential mechanism
is an inchwormlike process, similar to that reported in PAMPS gels [Osada and
Gong, 1993]. In that mechanism the cylindrical gel undergoes repeated cycles of
curling and straightening. The ends of the gel are hooked to a ratchet in such a
way that each bending cycle results in a step advance. Repeated cycling advances
the gel by a significant magnitude.
The same principle could apply in the actin filament. If the filament were to
pass through cycles of curling and straightening, such action could be harnessed
to propel the filament toward the center of the sarcomere. More realistically, any
such curling would arise from a local phase-change, which would then propagate
along the filament to produce a wormlike reptation. Each cycle of propagation
would result in a step of filament translation. The extent of translation would be a
function of the number of cycles. A schematic illustration of such an inchworm
process is shown in Fig. 12.
The wavelike motion that would be anticipated in the actin filament is
broadly observed [for review, cf. Pollack, 1996]. The evidence draws from as
early as four decades ago when undulations were directly observed to propagate
along actin-filament bundles responsible for active streaming. It also follows
from modern studies of single actin filaments. Such motion could be generated
by a local molecular structural change, which is observed in the electron
microscope [Menetret et al., 1991] and in actin crystals [Schutt and Lindberg,
1992]. That structural changes can propagate along the filament is supported by
the observation that the binding of a ligand to one end of the actin filament
affects the physical and mechanical properties of the entire filament
[Prochniewicz et al., 1996]. Indeed, direct microscopic visualization of actin
filaments shows a translation pattern quite strikingly characteristic of snakelike
motion [deBeer et al., 1997].

Figure 12: Reptation model explaining translation of thin filament over thick
filament. Black dots represent myosin cross-bridges. In this model a phase-
transition propagates from tail to head, advancing the thin filament by one notch.

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 Natural Muscle as a Biological System 67

Evidence for inchwormlike behavior is also inferred from dynamics of the

intact sarcomere, as shown below. During active sliding of thin filaments past
thick, the pattern of sarcomere shortening is stepwise. The measurement is made
by projecting the striated image of the myofibril onto a photodiode array, and
scanning the array regularly to track individual sarcomere length. Step sizes were
measured initially in Blyakhman et al. [1999], and an updated but similar result is
shown in the histogram of Fig. 13. The histogram shows multiple peaks,
indicating steps of discrete size. Although interpeak spacing is not precisely
uniform, there is a strong tendency for peaks to repeat at regular intervals. The
best-fit spacing between peaks is 2.7 nm, indicating a step size that is an integer
multiple of 2.7 nm. This value is equal to the linear advance of actin monomers
along the thin filament.
Such “quantal” behavior is inevitable if an actin monomer is bound to the
(immobile) cross-bridge between translation steps; filament translation steps
must then be n x the actin-monomer advance (Fig. 14). That actin and myosin
bind to one another is well recognized, and it is presumed that such binding is
responsible for the sustenance of active tension. Successive bindings give rise to
the inevitable quantum step. The size of the quantum, 2.7 nm, follows the
expectation of the inchwormlike mechanism.

Figure 13: Histogram of step size measured during active contraction of single
isolated myofibrils. Spacing between major peaks is 2.7 nm [Yakovenko et al.,
2002]. (Courtesy of the American Physiological Society.)

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 68 Chapter 2

Figure 14: Inchworm advance. The model predicts that the size of the step
advance will be an integer multiple of the actin monomer spacing.

Of particular interest is the fact that these active contraction steps are
preserved at higher levels of organization. Not only do the steps appear in the
single sarcomere, they appear as well at the level of the single cell [Pollack et al.,
1977]. If the process is considered as a phase transition, the transition is, again,
highly cooperative.

2.5 Lessons from the Natural Muscle System That May

Be Useful for the Design of Polymer Actuators
There is ample evidence that the filamentous polymers of the sarcomere—all
three filamentous polymers—shorten in a discrete, cooperative manner. If so,
muscle contraction very much resembles phase transitions that occur in synthetic
polymeric systems. The biological transition may, however, be a lot “smarter”
because of the sophistication of its responsiveness. Thus, the connecting filament
shortens in the absence of any activation; it behaves as a discrete elastic band.
The actin filament undergoes transition after the level of activation crosses
threshold. Because the actin filament, but not the myosin filament, is found in
relatively simple motile cells, the actin mechanism may be more primitive. Like
other primitive mechanisms, it is limited in its capacity and unable to function
under too high a load—just as a weight hung on a caterpillar’s tail can inhibit
upward crawling, even though the caterpillar may still cling. Beyond this critical
load, the operative agent is the thick filament, which can shorten under the
highest of loads. Indeed, the helix-coil transition has been demonstrated to
produce a force that accounts quantitatively for the maximum force muscle can

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 Natural Muscle as a Biological System 69

produce [Harrington, 1979]. Thus, the muscle polymer gel capitalizes on the
strengths of all three proteins. This is perhaps why muscle, a highly evolved
organelle, is as versatile as it is.
One upshot of these considerations is that the mechanism in muscle
contraction is not as different from the mechanisms used to construct artificial
muscles as had been thought. Both employ polymer-gel phase transitions. Like
the polymer gel, the muscle can be triggered by any variety of stimuli ranging
from a change of pH, salt, temperature, solvent type, and even electrical current.
Therefore, it may be possible to construct artificial muscles with greater speed
and efficacy by looking more closely at how nature accomplishes the task of
biological contraction.
This can be approached in several ways. One way is to use gels constructed
of polymers having some of the same features as muscle proteins. It may be
possible, for example, to construct simple gels of only one of the three protein
filaments. These polymers could be arranged randomly, or perhaps cross-linked
to one another along their length in a parallel arrangement. The parallel, cross-
linked arrangement may be particularly advantageous for achieving high speed:
cooperativity may be enhanced because the linked units must act in unison, or not
at all. Muscle can contract very rapidly. A common difference between natural
and artificial muscle is that natural muscle filaments (as well as other biological
filaments) are generally parallel and cross-linked, whereas artificial gels are
typically random. It may be that the parallel arrangement is a critical factor for
high speed.
Cross-linking may also be critical. Muscle filaments are hydrophilic and tend
to adsorb water. It is common experience that muscle water is difficult to
remove, even by centrifugation. In the absence of cross-linking, muscle-filament
polymers will swell, much like the gel of Fig. 6. Cross-links keep filaments
closely packed and unable to swell appreciably. The cross-linked matrix
therefore has more potential energy than the uncross-linked matrix, and may use
some of this chemical potential to contract more powerfully.
A related consideration is size. One conspicuous feature of muscle is that it is
built of many parallel units, or myofibrils (Fig. 1). Myofibril diameter is
consistently 1–2 μm, rarely more. This diminutive structure is surrounded by its
own activating system, called the sarcoplasmic reticulum, which supplies the
activating signal. Thus, diffusion distances are only 1–2 μm in natural muscle,
which presumably contributes to high speed. Emulation of this kind of
arrangement could result in higher contraction speed in artificial actuators.
As for the type of polymer that might be most effective, again, copying
nature’s design could lead to materials more responsive than those used to date.
For high force applications, the choice may be a myosinlike polymer. The
myosin melt force, as mentioned, is sufficient to account for full muscle tension.
For small-scale snakelike movements, the actin-based gel may be a better choice.
For bending, nature often employs microtubules, built of the protein tubulin. The
axostyle, for example, consists of a bundle of parallel microtubules cross-linked
along their length. Bending of the axostyle is frequently associated with

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 70 Chapter 2

shortening of microtubules on one edge relative to the other [McIntosh, 1973],

like a bimetal strip. The challenge is to synthesize polymers that have many of
the features of these proteins—or, to use the proteins themselves.
These are a few directions implied by the success of Mother Nature. It may
be that artificial actuators could ultimately produce movement comparable to
what nature has had almost four billion years to produce.

2.6 References
Baatsen, P. H. W. W., K. Trombitas, and G. H. Pollack, “Thick filaments of
striated muscle are laterally interconnected,” J. Ultras. & Mol. Str. Res. 97,
39-49, 1988.
Blyakhman, F., T. Shklyar, G. H. Pollack, “Quantal length changes in single
contracting sarcomeres,” J. Mus Res. Cell Motility, 20, 529-538, 1999.
Cooke, R., “Actomyosin interaction in striated muscle,” Physiol. Rev. 77(3): 671-
697, 1997.
deBeer, E. L., A. M. A T. A. Sontrop, M. S. Z. Kellermayer, G. H. Pollack,
“Actin-filament motion in the in vitro motility assay has a periodic
component,” Cell Motil. Cytoskel. 38, 341-50, 1997.
Granzier, H. L. M., and G. H. Pollack, “Stepwise shortening in unstimulated frog
skeletal muscle fibres,” J. Physiol. 362, 173-88, 1985.
Harrington, W. F., “On the origin of the contractile force in skeletal muscle,”
Proc. Nat’l. Acad. Sci. 76, 5066-70, 1979.
Hoffman, A. S., “Conventional and environmentally-sensitive hydrogels for
medical and industrial uses: a review paper,” Polymer Gels 268(5): 82-87,
1991.
Hoyle, G., Muscles and their Neural Control, Wiley, N. Y. 1983.
Huxley, A. F., “Muscle structure and theories of contraction,” Prog. Biophys.
Biophys. Chem. 7: 255-318, 1957.
Huxley, A. F., and R. Niedergerke, “Structural changes in muscle during
contraction: Interference microscopy of living muscle fibres,” Nature 173:
971-973, 1954.
Huxley, A. F., and R. Niedergerke, “Measurement of the striations of isolated
muscle fibres with the interference microscope,” J. Physiol. 144: 403-425,
1958.
Huxley, H. E., “A personal view of muscle and motility mechanisms,” Ann. Rev.
Physiol. 58: 1-19, 1996.
Huxley, H. E., and J. Hanson, “Changes in the cross-striations of muscle during
contraction and stretch and their structural interpretation,” Nature 173: 973-
976, 1954.
Huxley, H. E., and W. Brown, “The low angle X-ray diagram of vertebrate
striated muscle and its behaviour during contraction and rigor,” J. Mol. Biol.
39, 383-434, 1967.
Iwazumi, T, “A new field theory of muscle contraction,” Ph. D. Dissertation,
Univ. of Pennsylvania, 1970.

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 Natural Muscle as a Biological System 71

Katayama, E., “Quick-freeze deep-etch electron microscopy of the actin - heavy

meromyosin complex during the in vitro motility assay,” J. Mol. Biol. 278:
349-367, 1998.
McIntosh, J. R., “The axostyle of Saccinobaculus. II. Motion of the microtubule
bundle and a structural comparison of straight and bent axostyles,” J. Cell
Biol. 56: 324-339, 1973.
Menetret, J. F., W. Hoffman, R. R. Schroder, R. Gapp, R. S. Goody, “Time-
resolved cryo-electron microscopic study of the dissociation of actomyosin
induced by photolysis of photolabile nucleotides,” J. Mol. Biol. 219, 139-44,
1991.
Oplatka, A., “The rise, decline, and fall of the swinging crossbridge dogma,”
Chemtracts Bioch. Mol. Biol. 6, 18-60, 1996.
Oplatka, A, “Critical review of the swinging cross-bridge theory and of the
cardinal active role of water in muscle contraction,” Crit. Rev. Biochem. Mol.
Biol. 32, 307-60, 1997.
Osada, Y. and J. Gong, “Stimuli-responsive polymer gels and their application to
chemomechanical systems,” Prog. Polym. Sci. 18, 187, 1993.
Pollack, G. H., T. Iwazumi, H. E. D. J. ter Keurs, and E. F. Shibata, “Sarcomere
shortening in striated muscle occurs in stepwise fashion,” Nature 268, 757-9,
1977.
Pollack, Gerald H., Cells, Gels, and the Engines of Life, Ebner and Sons, Seattle,
2001.
Pollack, G. H., Muscle and Molecules: Uncovering the Principles of Biological
Motion, Ebner and Sons, Seattle, 1990.
Pollack, G. H., “Phase transitions and the molecular mechanism of contraction,”
Biophys. Chem. 59, 315-28, 1996.
Pollack, G. H., “The cross-bridge theory,” Physiol. Reviews 63, 1049-113, 1983.
Prochniewicz, E., Q. Zhang, P. A. Janmey, D. D. Thomas, “Cooperativity in F-
actin: Binding of gelsolin at the barbed end affects structure and dynamics of
the whole filament,” J. Mol. Biol. 260, 756-66, 1996.
Rief, M., M. Gautel, F. Oesterhelt, J. M. Fernandez, and H. E. Gaub, “Reversible
unfolding of individual titin immunoglobulin domains by AFM,” Science
276, 1109-12, 1997.
Schutt, C. E. and U. Lindberg, “A new perspective on muscle contraction,” FEBS
Lett. 325: 59-62, 1993.
Schutt, C. E. and U. Lindberg, “Actin as a generator of tension during muscle
contraction,” Proc. Nat’l. Acad. Sci. 89, 319-23, 1992.
Schutt, C. E. and U. Lindberg, “Muscle contraction as a Markov process
I:enertetics of the process,” Acta Physiol. Scan. 163: 307-324, 1998.
Spudich, J. A., “How molecular motors work,” Nature 372: 515-518, 1994.
Tanaka, T., “Gels,” Sci. Amer. 244, 110, 1981.
Thomas, D. D, “Spectroscopic probes of muscle cross-bridge rotation,” Ann Rev.
Physiol. 49: 641-709, 1987.
Trombitas, K. P. H. W. W. Baatsen, and G. H. Pollack, “I-bands of striated
muscle contain lateral struts,” J. Ultras. & Mol. Str. Res. 100, 13-30, 1988.

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 72 Chapter 2

Tskhovrebova, L., J. Trinick, J. A. Sleep, and R. M. Simmons, “Elasticity and

unfolding of single molucules of the giant muscle protein titin,” Nature 387,
308-12, 1997.
Yagi, N. and I. Matsubara, “Cross-bridge movements during slow length change
of active muscle,” Biophys. J. 45, 611-4, 1984.
Yakovenko, O., Blyakhman, F. and Pollack, G. H., “Fundamental step size in
single cardiac and skeletal sarcomeres,” Am J. Physiol (Cell) 283(3): C735-
C743, 2002.
Yang, P., T. Tameyasu, and G. H. Pollack, “Stepwise dynamics of connecting
filaments measured in single myofibrillar sarcomeres,” Biophys. J. 74, 1473-
83, 1998.

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