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M Sokolov

Publications and source records attributed to M Sokolov.

5 recordsLinked to original sources

The 20 C-terminal amino acid residues of the chloroplast ATP synthase gamma subunit are not essential for activity.

It has been suggested that the last seven to nine amino acid residues at the C terminus of the gamma subunit of the ATP synthase act as a spindle for rotation of the gamma subunit with respect to the alpha beta subunits during catalysis (Abrahams, J. P., Leslie, A. G. W., Lutter, R., and Walker, J. E. (1994) Nature 370, 621-628). To test this hypothesis we selectively deleted C-terminal residues from the chloroplast gamma subunit, two at a time starting at the sixth residue from the end and finishing at the 20th residue from the end. The mutant gamma genes were overexpressed in Escherichia coli and assembled with a native alpha3beta3 complex. All the mutant forms of gamma assembled as effectively as the wild-type gamma. Deletion of the terminal 6 residues of gamma resulted in a significant increase (>50%) in the Ca-dependent ATPase activity when compared with the wild-type assembly. The increased activity persisted even after deletion of the C-terminal 14 residues, well beyond the seven residues proposed to form the spindle. Further deletions resulted in a decreased activity to approximately 19% of that of the wild-type enzyme after deleting all 20 C-terminal residues. The results indicate that the tip of the gammaC terminus is not essential for catalysis and raise questions about the role of the C terminus as a spindle for rotation.

Amino Acid Sequence↗

Evidence for an ephaptic feedback in cortical synapses: postsynaptic hyperpolarization alters the number of response failures and quantal content.

The amplitude of excitatory postsynaptic potentials and currents increases with membrane potential hyperpolarization. This has been attributed to an increase in the driving force when the membrane potential deviates from the equilibrium potential of the respective ions. Here we report that in a subset of neocortical and hippocampal synapses, postsynaptic hyperpolarization affects traditional measures of transmitter release: the number of failures, coefficient of variation of response amplitudes, and quantal content, suggesting increased presynaptic release. The result is compatible with the hypothesis of Byzov on the existence of electrical (or "ephaptic") linking in purely chemical synapses. The linking, although negligible at neuromuscular junctions, could be functionally significant in influencing transmitter release at synapses with high resistance along the synaptic cleft. Our findings necessitate reconsideration of classical amplitude-voltage relations for such synapses. Thus, synaptic strength may be enhanced by hyperpolarization of the postsynaptic membrane potential. The positive ephaptic feedback could account for "all-or-none" excitatory postsynaptic potentials at some cortical synapses, large evoked and spontaneous multiquantal events and a high efficacy of large "perforated" synapses whose number increases following behavioural learning or the induction of long-term potentiation.

2-Amino-5-phosphonovalerate↗

Spinach chloroplast coupling factor CF1-alpha 3 beta 3 core complex: structure, stability, and catalytic properties.

A minimal chloroplast coupling factor CF1 core complex, containing only alpha and beta subunits, has been isolated from spinach thylakoids [Avital, S., & Gromet-Elhanan, Z. (1991) J. Biol. Chem. 266, 7067-7072]. This CF1(alpha beta) exhibited a low MgATPase activity, which was stimulated but not inhibited by low concentrations of the species-specific CF1 effector tentoxin. As is reported here, the structure of CF1(alpha beta) could not be determined due to its instability. However, its pretreatment with high tentoxin concentrations resulted in a remarkable 50-fold stimulation of the MgATPase activity as well as stabilization of its hexameric structure, thus enabling the isolation of a more active CF1-alpha 3 beta 3 complex by size-exclusion chromatography. A detailed characterization of the MgATPase activity of this tentoxin-stabilized CF1-alpha 3 beta 3 hexamer, as compared to the activity of a CF1 complex lacking the epsilon subunit, revealed similar apparent Km values and a similar stimulation by the presence of 100 microM tentoxin in the assay medium, but drastic differences in all other tested assays. Most pronounced were their different temperature profiles and different responses to all added inhibitors and stimulators of the CF1 MgATPase activity and to excess free Mg2+ ions. The specific properties of the stable CF1-alpha 3 beta 3 hexamer are identical to those earlier reported for its parent-unstable CF1(alpha beta). These results indicate that, although the CF1 gamma subunit is not required for the low CF1(alpha beta) ATPase activity nor for the higher activity of the tentoxin-stabilized CF1-alpha 3 beta 3, it plays a central role in obtaining the typical functional properties of the CF1-ATPase. Kinetic cooperativity could not be critically tested as yet with any F1-alpha 3 beta 3. However, tentoxin, as azide, has been shown to inhibit multisite but not unisite catalysis. Therefore, the observation that CF1-alpha 3 beta 3 is only stimulated by tentoxin suggests that the required presence of CF1-gamma for obtaining inhibition by tentoxin reflects the role of this subunit in cooperative interactions between the catalytic sites.

Adenosine Triphosphate↗

An experimental study of pulsatile pipe flow in the transition range.

The study of pulsatile flows is relevant to many areas of applications. Typical applications include aerodynamics, biofluid mechanics, wind flows, and gas transport. Transition to turbulence during pulsatile flow is physiologically and clinically important. It has been suggested as a possible mechanism to enhance the transport of gases during high-frequency ventilation, may be related to valvular regurgitation and heart murmurs and to post stenotic dilatation and aneurysms. Measurements in a pulsatile pipe flow with a superimposed mean flow are reported. Data were taken in a water flow with mean Reynolds numbers in the range of 0 < Re(m) < 3000, oscillating Reynolds numbers of 0 < Re omega < 4000, and Stokes parameter 7 < lambda < 15. Velocity profiles of various phases of the flow, condition for flow reversal, and pressure losses were measured. The adequacy of a quasi-steady-state model is discussed. Condition for transition is determined by visually inspecting velocity signals at the centerline.

Biomechanical Phenomena↗