Biomedical subjects

O A Andreev

Publications and source records attributed to O A Andreev.

3 recordsLinked to original sources

PubMed1992

Two different acto-S1 complexes.

Based on change in anisotropy of fluorescently labelled S1 and on increase in turbidity of acto-S1 complex when S1 bound to F-actin, we reported previously that depending on the molar ratio of S1 to actin two different complexes of actin monomer (A) and myosin subfragment 1 (S1) could be formed: A1*S1 (one actin with one S1) and A2*S1 (two actins with one S1). Here we extend these findings to F-actin labelled with pyrene and cross-linked to S1 with 1-ethyl-3-(3-dimethyl-aminopropyl)carbodiimide (EDC). The fluorescence of pyrene F-actin decreased with increase in S1 concentration and reached saturation at a molar ratio of S1 to actin of either 0.5 or 1.0, depending on whether S1 was added slowly (5 min) or quickly (10-20 s between additions). Incubation of A2*S1 complex in excess of S1 for > 1 h caused a shift in equilibrium towards the A1*S1 complex. The A2*S1 complexes were not formed at high S1 to actin ratios (> 1.0) owing to competition between heads. Crosslinking experiments showed that the formation of EDC crosslinked products, 175-185 kDa doublet and 265 kDa band, depended on the ratio S1 to actin. To assess the relative ratio of S1 and actin in crosslinked products, we labelled S1 and F-actin with different fluorescent probes (5-IAF and IATR). The S1 to actin ratio was proportional to the ratio of intensities of fluorescence of labelled S1 and actin. The S1 to actin ratio in 265 kDa product was two times smaller than in 175-185 kDa doublet (which is believed to be A1*S1 complex) and therefore 265 kDa band corresponded to A2*S1. Transition between two types of binding may be important to understanding how muscle contracts.

Actin Cytoskeleton↗

PubMed1991

The myosin head can bind two actin monomers.

Force impulse is thought to be generated in muscle when myosin head (S-1), while weakly bound to actin filament, undergoes orientational change to form a strong (rigor) bond with actin. There is ample evidence that this bond involves interaction of 1 myosin head with 1 actin monomer. However, X-ray diffraction data of muscle decorated with S-1, as well as recently proposed model of the thin filaments, suggested that each S-1 molecule interacted with two actin monomers. We reinvestigated this controversy and found that the stoichiometry of acto-S-1 bond depended on the relative amounts of actin and myosin present during titrations: when increasing amounts of actin were added to a fixed amount of S-1 (i.e. when myosin heads were initially in excess over actin), the saturating stoichiometry was 1 mol of S-1 per 1 mol of actin. However, when increasing amounts of S-1 were added slowly to a fixed amount of F-actin (i.e. when actin was initially in excess over S-1), the stoichiometry at saturation was 1 mol of S-1 per 2 mols of actin. The ability of S-1 to bind either one or two actin monomers suggests a way that force could be generated during muscle contraction.

Actins↗

PubMedDate not supplied

[Effect of polyamines on the thermal resistance of Escherichia coli cells in heat shock].

Polyamines intensify the effect of a heat shock on Escherichia coli M-17 cells. The lethal effect of polyamines rises in the spermidine--spermine series as their concentration and the duration of heat action is increased. The effect of polyamines on the cells subjected to a heat shock is not associated with the activity of amine oxidases and, apparently, does not destabilize the membranes. As was demonstrated using electron microscopy, the cells undergo morphological changes in the presence of polyamines in the course of a heat shock; in particular, electron-dense regions appear in the nucleoid zone, presumably, due to DNA conformational rearrangements during the shock in the presence of polyamines.

Cell Nucleus↗