Muscle contraction is one of those biological phenomena that we can all appreciate in our everyda... more Muscle contraction is one of those biological phenomena that we can all appreciate in our everyday lives. Sometimes it is when we are resting quietly and are aware of our heartbeat. At other times it may be when we are exerting ourselves and become short of breath, or when we exercise for a long period and our muscles start to ache. The way in which muscles produce force has exercised the minds of philosophers and scientists at least since the days of Erasistratus in the third century BC. Nowadays, of course, we know a very great deal about muscle structure, physiology and biochemistry, but we still do not know exactly what the molecular process is that produces movement. An ideal way of probing this process would be to be able to obtain signals from the relevant molecules as they actually go through their normal force-generating routine in an active muscle. The spatial dimensions involved are in the region of 1 - 50 nm, thus precluding the use of light microscopy, and the time regime is microseconds to milliseconds. Techniques with the appropriate spatial resolution might be electron microscopy and x-ray diffraction, but electron microscopy cannot yet be carried out on living tissue. X-ray diffraction methods can clearly have the right sort of spatial resolution, but what about recording diffraction patterns in the very short times involved (say 1 ms)? It is here that the high flux from synchrotron storage rings comes into its own. Using synchrotron radiation from, say, the SRS at the CCLRC Daresbury Laboratory it is possible to record x-ray diffraction patterns from living muscles in the millisecond time regime and to follow how these diffraction patterns change as the muscles go through typical contraction cycles. Unfortunately, x-ray diffraction is not a direct imaging method; the observed distribution of diffracted intensity needs to be interpreted in some way to give useful information on the spatial relationships of the force-generating molecules. This review details the practical methods involved in recording time-resolved x-ray diffraction patterns from active muscles and the theoretical approaches that are being used to interpret the diffraction patterns that are obtained. The ultimate aim is to produce a series of time-sliced images of the changing molecular arrangements and shapes in the muscle as force is produced; together these images will form `Muscle - The Movie'.
It is 30 years since Ebashi and colleagues showed that Ca 2/ ions directly affect regulation of t... more It is 30 years since Ebashi and colleagues showed that Ca 2/ ions directly affect regulation of the myosin-actin interaction in muscle through the action of tropomyosin and troponin on muscle thin filaments. It is more than 20 years since the idea was put forward that tropomyosin might act, at least in part, by changing its position on actin, thus uncovering or modifying the myosin binding site on actin when troponin molecules take up Ca 2/ . Since that time, a great deal of evidence for and against this steric blocking mechanism has been published: a structure for actin filaments at close to atomic resolution has been proposed, and the whole regulation story has become both more complicated and more subtle. Here we review structural and biochemical aspects of regulation in vertebrate skeletal muscle. We show that some basic ideas of the steric blocking mechanism remain valid. We also show that additional factors, such as troponin movements and structural changes within the actin monomers themselves, may be crucial. A number of the resulting regulation scenarios need to be distinguished.-Squire, J. M., Morris, E. P. A new look at thin filament regulation in vertebrate skeletal muscle. FASEB J. 12, 761-771 (1998)
Myosin filaments in muscle, carrying the ATPase myosin heads that interact with actin filaments t... more Myosin filaments in muscle, carrying the ATPase myosin heads that interact with actin filaments to produce force and movement, come in multiple varieties depending on species and functional need, but most are based on a common structural theme. The now successful journeys to solve the ultrastructures of many of these myosin filaments, at least at modest resolution, have not been without their false starts and erroneous sidetracks, but the picture now emerging is of both diversity in the rotational symmetries of different filaments and a degree of commonality in the way the myosin heads are organised in resting muscle. Some of the remaining differences may be associated with how the muscle is regulated. Several proteins in cardiac muscle myosin filaments can carry mutations associated with heart disease, so the elucidation of myosin filament structure to understand the effects of these mutations has a clear and topical clinical relevance.
We report on recent developments in CCP13 software with particular emphasis on ICE, an integrated... more We report on recent developments in CCP13 software with particular emphasis on ICE, an integrated, internet-enabled, common desktop CCP13 environment. As an initial package, several heavily used data conversion and stripping programs have been integrated together in ICE with an easy to use Graphical User Interface (GUI) and enhanced functionality. ICE can dynamically group CCP13 programs together by using a
Myosin filaments in muscle, carrying the ATPase myosin heads that interact with actin filaments t... more Myosin filaments in muscle, carrying the ATPase myosin heads that interact with actin filaments to produce force and movement, come in multiple varieties depending on species and functional need, but most are based on a common structural theme. The now successful journeys to solve the ultrastructures of many of these myosin filaments, at least at modest resolution, have not been without their false starts and erroneous sidetracks, but the picture now emerging is of both diversity in the rotational symmetries of different filaments and a degree of commonality in the way the myosin heads are organised in resting muscle. Some of the remaining differences may be associated with how the muscle is regulated. Several proteins in cardiac muscle myosin filaments can carry mutations associated with heart disease, so the elucidation of myosin filament structure to understand the effects of these mutations has a clear and topical clinical relevance.
Myosin filaments in vertebrate striated muscle have a long roughly cylindrical backbone with cros... more Myosin filaments in vertebrate striated muscle have a long roughly cylindrical backbone with cross-bridge projections on the surfaces of both halves except for a short central bare zone. In the middle of this central region the filaments are cross-linked by the M-band which holds them in a well-defined hexagonal lattice in the muscle A-band. During muscular contraction the M-band-defined rotation of the myosin filaments around their long axes influences the interactions that the cross-bridges can make with the neighbouring actin filaments. We can visualise this filament rotation by electron microscopy of thin cross-sections in the bare-region immediately adjacent to the M-band where the filament profiles are distinctly triangular. In the muscles of teleost fishes, the thick filament triangular profiles have a single orientation giving what we call the simple lattice. In other vertebrates, for example all the tetrapods, the thick filaments have one of two orientations where the trian...
FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 1998
It is 30 years since Ebashi and colleagues showed that Ca2+ ions directly affect regulation of th... more It is 30 years since Ebashi and colleagues showed that Ca2+ ions directly affect regulation of the myosin-actin interaction in muscle through the action of tropomyosin and troponin on muscle thin filaments. It is more than 20 years since the idea was put forward that tropomyosin might act, at least in part, by changing its position on actin, thus uncovering or modifying the myosin binding site on actin when troponin molecules take up Ca2+. Since that time, a great deal of evidence for and against this steric blocking mechanism has been published: a structure for actin filaments at close to atomic resolution has been proposed, and the whole regulation story has become both more complicated and more subtle. Here we review structural and biochemical aspects of regulation in vertebrate skeletal muscle. We show that some basic ideas of the steric blocking mechanism remain valid. We also show that additional factors, such as troponin movements and structural changes within the actin monom...
Advances in experimental medicine and biology, 1993
The advantages of using time-resolved X-ray diffraction as a means of probing myosin cross-bridge... more The advantages of using time-resolved X-ray diffraction as a means of probing myosin cross-bridge behaviour in active muscle are outlined, together with the reasons that bony fish muscle has advantages in such studies. We show that the observed X-ray diffraction patterns from fish muscle can be analysed in a way that is rigorous enough to allow reliable information about crossbridge activity to be defined. Among the advantages of this muscle are that diffraction patterns from resting, active and rigor muscles are all well-sampled at least out to the 30 row-line, that the resting myosin layer-line pattern can be 'solved' crystallographically to define the starting position of the crossbridges in resting muscle, and that the equatorial intensity distribution, which in all patterns from vertebrate skeletal muscles comprises overlapping peaks from the A-band and the Z-band, can be analysed sufficiently rigorously to allow separation of the two patterns, both of which change when...
Muscle contraction is one of those biological phenomena that we can all appreciate in our everyda... more Muscle contraction is one of those biological phenomena that we can all appreciate in our everyday lives. Sometimes it is when we are resting quietly and are aware of our heartbeat. At other times it may be when we are exerting ourselves and become short of breath, or when we exercise for a long period and our muscles start to ache. The way in which muscles produce force has exercised the minds of philosophers and scientists at least since the days of Erasistratus in the third century BC. Nowadays, of course, we know a very great deal about muscle structure, physiology and biochemistry, but we still do not know exactly what the molecular process is that produces movement. An ideal way of probing this process would be to be able to obtain signals from the relevant molecules as they actually go through their normal force-generating routine in an active muscle. The spatial dimensions involved are in the region of 1 - 50 nm, thus precluding the use of light microscopy, and the time regime is microseconds to milliseconds. Techniques with the appropriate spatial resolution might be electron microscopy and x-ray diffraction, but electron microscopy cannot yet be carried out on living tissue. X-ray diffraction methods can clearly have the right sort of spatial resolution, but what about recording diffraction patterns in the very short times involved (say 1 ms)? It is here that the high flux from synchrotron storage rings comes into its own. Using synchrotron radiation from, say, the SRS at the CCLRC Daresbury Laboratory it is possible to record x-ray diffraction patterns from living muscles in the millisecond time regime and to follow how these diffraction patterns change as the muscles go through typical contraction cycles. Unfortunately, x-ray diffraction is not a direct imaging method; the observed distribution of diffracted intensity needs to be interpreted in some way to give useful information on the spatial relationships of the force-generating molecules. This review details the practical methods involved in recording time-resolved x-ray diffraction patterns from active muscles and the theoretical approaches that are being used to interpret the diffraction patterns that are obtained. The ultimate aim is to produce a series of time-sliced images of the changing molecular arrangements and shapes in the muscle as force is produced; together these images will form `Muscle - The Movie'.
It is 30 years since Ebashi and colleagues showed that Ca 2/ ions directly affect regulation of t... more It is 30 years since Ebashi and colleagues showed that Ca 2/ ions directly affect regulation of the myosin-actin interaction in muscle through the action of tropomyosin and troponin on muscle thin filaments. It is more than 20 years since the idea was put forward that tropomyosin might act, at least in part, by changing its position on actin, thus uncovering or modifying the myosin binding site on actin when troponin molecules take up Ca 2/ . Since that time, a great deal of evidence for and against this steric blocking mechanism has been published: a structure for actin filaments at close to atomic resolution has been proposed, and the whole regulation story has become both more complicated and more subtle. Here we review structural and biochemical aspects of regulation in vertebrate skeletal muscle. We show that some basic ideas of the steric blocking mechanism remain valid. We also show that additional factors, such as troponin movements and structural changes within the actin monomers themselves, may be crucial. A number of the resulting regulation scenarios need to be distinguished.-Squire, J. M., Morris, E. P. A new look at thin filament regulation in vertebrate skeletal muscle. FASEB J. 12, 761-771 (1998)
Myosin filaments in muscle, carrying the ATPase myosin heads that interact with actin filaments t... more Myosin filaments in muscle, carrying the ATPase myosin heads that interact with actin filaments to produce force and movement, come in multiple varieties depending on species and functional need, but most are based on a common structural theme. The now successful journeys to solve the ultrastructures of many of these myosin filaments, at least at modest resolution, have not been without their false starts and erroneous sidetracks, but the picture now emerging is of both diversity in the rotational symmetries of different filaments and a degree of commonality in the way the myosin heads are organised in resting muscle. Some of the remaining differences may be associated with how the muscle is regulated. Several proteins in cardiac muscle myosin filaments can carry mutations associated with heart disease, so the elucidation of myosin filament structure to understand the effects of these mutations has a clear and topical clinical relevance.
We report on recent developments in CCP13 software with particular emphasis on ICE, an integrated... more We report on recent developments in CCP13 software with particular emphasis on ICE, an integrated, internet-enabled, common desktop CCP13 environment. As an initial package, several heavily used data conversion and stripping programs have been integrated together in ICE with an easy to use Graphical User Interface (GUI) and enhanced functionality. ICE can dynamically group CCP13 programs together by using a
Myosin filaments in muscle, carrying the ATPase myosin heads that interact with actin filaments t... more Myosin filaments in muscle, carrying the ATPase myosin heads that interact with actin filaments to produce force and movement, come in multiple varieties depending on species and functional need, but most are based on a common structural theme. The now successful journeys to solve the ultrastructures of many of these myosin filaments, at least at modest resolution, have not been without their false starts and erroneous sidetracks, but the picture now emerging is of both diversity in the rotational symmetries of different filaments and a degree of commonality in the way the myosin heads are organised in resting muscle. Some of the remaining differences may be associated with how the muscle is regulated. Several proteins in cardiac muscle myosin filaments can carry mutations associated with heart disease, so the elucidation of myosin filament structure to understand the effects of these mutations has a clear and topical clinical relevance.
Myosin filaments in vertebrate striated muscle have a long roughly cylindrical backbone with cros... more Myosin filaments in vertebrate striated muscle have a long roughly cylindrical backbone with cross-bridge projections on the surfaces of both halves except for a short central bare zone. In the middle of this central region the filaments are cross-linked by the M-band which holds them in a well-defined hexagonal lattice in the muscle A-band. During muscular contraction the M-band-defined rotation of the myosin filaments around their long axes influences the interactions that the cross-bridges can make with the neighbouring actin filaments. We can visualise this filament rotation by electron microscopy of thin cross-sections in the bare-region immediately adjacent to the M-band where the filament profiles are distinctly triangular. In the muscles of teleost fishes, the thick filament triangular profiles have a single orientation giving what we call the simple lattice. In other vertebrates, for example all the tetrapods, the thick filaments have one of two orientations where the trian...
FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 1998
It is 30 years since Ebashi and colleagues showed that Ca2+ ions directly affect regulation of th... more It is 30 years since Ebashi and colleagues showed that Ca2+ ions directly affect regulation of the myosin-actin interaction in muscle through the action of tropomyosin and troponin on muscle thin filaments. It is more than 20 years since the idea was put forward that tropomyosin might act, at least in part, by changing its position on actin, thus uncovering or modifying the myosin binding site on actin when troponin molecules take up Ca2+. Since that time, a great deal of evidence for and against this steric blocking mechanism has been published: a structure for actin filaments at close to atomic resolution has been proposed, and the whole regulation story has become both more complicated and more subtle. Here we review structural and biochemical aspects of regulation in vertebrate skeletal muscle. We show that some basic ideas of the steric blocking mechanism remain valid. We also show that additional factors, such as troponin movements and structural changes within the actin monom...
Advances in experimental medicine and biology, 1993
The advantages of using time-resolved X-ray diffraction as a means of probing myosin cross-bridge... more The advantages of using time-resolved X-ray diffraction as a means of probing myosin cross-bridge behaviour in active muscle are outlined, together with the reasons that bony fish muscle has advantages in such studies. We show that the observed X-ray diffraction patterns from fish muscle can be analysed in a way that is rigorous enough to allow reliable information about crossbridge activity to be defined. Among the advantages of this muscle are that diffraction patterns from resting, active and rigor muscles are all well-sampled at least out to the 30 row-line, that the resting myosin layer-line pattern can be 'solved' crystallographically to define the starting position of the crossbridges in resting muscle, and that the equatorial intensity distribution, which in all patterns from vertebrate skeletal muscles comprises overlapping peaks from the A-band and the Z-band, can be analysed sufficiently rigorously to allow separation of the two patterns, both of which change when...
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