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Biomedical subjects

R T Tregear

Publications and source records attributed to R T Tregear.

At least 37 records · Page 2Linked to original sources
PubMed1984

The repeat distance of myosin in the thick filaments of various muscles.

The meridional spacing of the X-ray diffraction peak from the repeat of myosin along the thick filament of four muscles has been remeasured on the same apparatus. The frog sartorius gave a shorter repeat distance (143.7 A) than the three invertebrate muscles, which ranged from 144.9 to 145.4 A. These results confirm earlier measurements. Provided that the myosin molecules are staggered relative to one another by a constant 98 residues, it may be inferred that in vertebrate thick filaments part or all of the tail lies at a considerable angle to the filament axis, whereas in the invertebrates the angle is smaller.

Animals↗
PubMed1984

Modification of the interactions of myosin with actin and 5'-adenylyl imidodiphosphate by substitution of ethylene glycol for water.

We studied the effect of replacing water by ethylene glycol as solvent on the properties of skeletal muscle myosin, myosin subfragment-1 (S1) and heavy meromyosin. Ethylene glycol (50%, v/v) had no detectable effect on the affinity of myosin or actomyosin for the substrate analogue 5'-adenylyl imidodiphosphate (AMPPNP). However, the rate constants for formation and dissociation of the myosin X MgAMPPNP complex were reduced 200-fold; the logarithm of the dissociation rate was roughly proportional to the fractional concentration of ethylene glycol. Nucleotide dissociation was accelerated at least 300-fold by pure actin but remained slow with regulated actin in the absence of Ca2+. Ethylene glycol substitution reduced the affinity of S1 and the S1 X MgAMPPNP complex for actin equally (100-fold at 50% ethylene glycol). These results show that ethylene glycol has specific effects on myosin's enzymic mechanism, which can account for its effect on the tension and stiffness of glycerinated muscle fibres.

Actins↗
PubMed1984

On the possibility of interaction between neighbouring crossbridges.

Demembranated insect or rabbit striated muscle fibres at equilibrium (i.e. in the absence of ATP hydrolysis) were modified either by substituting ethylene glycol for water or by adding AMPPNP. The resultant states observed by electron microscopy and X-ray diffraction appeared to contain a mixture of at least two distinct types of crossbridge, which were not randomly mixed. The crossbridges held tension for a great deal longer than they remained attached to actin in solution. In the presence of AMPPNP the muscle fibres relaxed at a critical glycol concentration. These properties indicate that the crossbridges interacted with one another.

Actins↗
PubMed1984

Modification of crossbridge states by ethylene glycol in insect flight muscle.

Substitution of ethylene glycol for part of the solvent water changes the mechanical properties, structure and nucleotide binding of glycerol-extracted flight muscle fibres from the waterbug Lethocerus. On addition of ethylene glycol the rigor tension falls, rapidly and reversibly. With increasing glycol concentration the effect saturates at a non-zero tension. The isotonic stiffness is unchanged on adding ethylene glycol. Adding MgAMPPNP (adenylylimidodiphosphate) to a muscle fibre in 50% ethylene glycol causes a further rapid tension fall; above 100 microM AMPPNP the tension reaches zero. The isotonic stiffness of restretched muscle is then close to that of a relaxed fibre. Removal of MgAMPPNP from the bathing medium has no immediate mechanical effect. After several hours the isotonic stiffness rises to some extent; on removal of the glycol both tension and stiffness rise to rigor values within one minute. 3H-Labelled AMPPNP binds to muscle fibres in 50% ethylene glycol in a similar amount to the number of myosin heads present. The binding is tighter than that in aqueous solution and the nucleotide is only released very slowly. Upon removal of the ethylene glycol nucleotide is rapidly released. X-ray diffraction of muscle in 50% ethylene glycol reveals a highly ordered structure, in which both the 14 nm and the 38 nm layer lines are sharply sampled and are of intermediate values between rigor and relaxation. The two inner equatorial peaks are also of intermediate values. On adding MgAMPPNP the pattern resembles that of relaxed muscle. Upon removal of the nucleotide the pattern does not revert towards rigor but on removal of glycol it does. These results are interpreted in terms of changes within the myosin heads and their array within the filament lattice.

Adenine Nucleotides↗
PubMed1984

The process of muscle relaxation by the combined action of MgAMPPNP and ethylene glycol.

Insect flight muscle fibres were relaxed by the combined action of MgAMPPNP and ethylene glycol, as measured by the stiffness of the fibres. Relaxation occurred over a small range of glycol concentration. Addition of Ca2+ raised the glycol required for relaxation. The speed at which the stiffness measurement was made did not influence the glycol concentration at which relaxation occurred. Glycol in excess of that needed to relax the muscle caused a slight rise in high-frequency stiffness. Removal of the glycol restored the rigor stiffness. Under glycol-relaxed conditions, much of the AMPPNP bound in muscle fibres was retained during cold-chase (elution of [3H] AMPPNP by nonradioactive AMPPNP); the intensity ratio of the inner equatorial X-ray diffraction peaks rose upon glycol relaxation to a value slightly below that characteristic of natural relaxation. The results are interpreted in terms of cooperative attachment of the crossbridges to actin.

Adenosine Triphosphate↗
PubMed1983

Generation of tension by glycerol-extracted vertebrate skeletal muscle fibres in the absence of calcium.

When a small bundle of glycerol-extracted fibres from either frog, tortoise or rabbit skeletal muscle was first exposed to high MgATP (5 mM) in the absence of Ca2+ (less than 1 nM) and at low ionic strength (less than 0.11) at 20 degrees C, it produced a single sharp transient contraction followed by a lower maintained isometric tension. The maintained tension was investigated further in rabbit psoas fibres. Ca2+-free tension was dependent on the ionic strength in the range 0.04-0.10, on the temperature in the range 6-20 degrees C and the free Mg2+ in the range 0-6 mM. It was promoted by low ionic strength, low Mg2+ and high temperature, and was unaffected by varying the MgATP2- in the range 0.4-4 mM and by adding ATP regenerating components. A separate regime of tension generation was detected at MgATP2- concentrations of less than 0.1 mM, in which MgATP2- concentration was critical. The results are interpreted on the assumption that binding of Mg2+ to some component of the regulatory system is necessary to maintain its inhibitory effect in the absence of Ca2+. Ionic strength and temperature, on the other hand, may affect actomyosin directly.

Adenosine Triphosphatases↗
PubMed1975

The relationship of adenosine triphosphatase activity to tension and power output of insect flight muscle.

1. On a simple model of actomyosin interaction, the tension cost (ATP hydrolysed/unit tension) and the frequency of low amplitude oscillation optimum for work production are both determined by the rate of detachment from the actin filament of the myosin crossbridge. To test this model, the two parameters were measured under different conditions using glycerol-extracted Lethocerus cordofanus dorsal longitudinal flight muscle fibres. 2. The ATPase activity of the static muscle rose by an amount approximately proportional to the rise in tension as the muscle was stretched. 3. When the muscle fibres were sinusoidally oscillated at 5-10 Hz by 2% of their resting length they produced a large amount of mechanical power and hydrolysed approximately twice as much ATP per unit mean tension as they did when static. The ATPase activity was linearly related to the mean tension during oscillation. 4. The experiments were repeated at temperatures between 12 and 30 degrees C and the tension cost and the optimal frequency of oscillation of the fibres were found to rise with temperature. 5. Removal of phosphate from the incubating medium reduced both the tension cost and the optimal working frequency. Addition of pyrophosphate or sulphate reduced both parameters still further. 6. From these results the tension cost of static muscle was shown to be proportional to its optimal working frequency. 7. ATPase activity rose monotonically with power production at work-producing frequencies and at moderate degrees of stretch. A high absolute efficiency was found under a wide range of conditions. 8. The proportionality between tension cost and optimal frequency is evidence for the proposed model of actomyosin interaction.

Adenosine Triphosphatases↗