Intracellular protein kinase and calcium inward currents in perfused neurones of the snail Helix pomatia.

Changes in the amplitude of the calcium inward current caused by intracellular administration of tolbutamide (an inhibitor of the cyclic AMP-dependent protein kinase activity) or catalytic subunits of cAMP-dependent protein kinases from rabbit myocardium were studied on internally perfused nerve cells of the snail, Helix pomatia. Intracellular administration of 7 mM tolbutamide caused a rapid decline of the amplitude of the calcium current that had been stabilized by theophylline; the effect was practically completely reversible. In contrast, addition to the perfusing solution of exogenous catalytic subunits of cyclic AMP-dependent protein kinases (about 0.7 microM of protein) together with 2 mM adenosine 5'-triphosphate and 3 mM MgCl2, led to stabilization of the calcium conductance of the cell membrane or restored it if it had declined during the perfusion with basic solution. The effect depended largely on the presence of adenosine 5'-triphosphate. Its time course was very slow (dozens of minutes) due probably to slow diffusion of the protein inside the cell. Heat-inactivated catalytic subunits did not produce such a stabilizing or restoring action on the calcium conductance. The results substantiate the suggestion that the normal functioning of calcium channels depends on phosphorylation catalyzed by cyclic AMP-dependent protein kinases.

[Heterogeneous and kinetic analysis of certain properties of Ca2+-ATPase in sarcolemma of rabbit skeletal muscles].

Kinetic properties of Ca2+-ATPase were studied by means of the continuous potentiometric method. On the basis of heterogenous and kinetic ideas it is shown that the experimental data on the effect of ions CA2+ on the Ca2+-ATPase activity may be explained, assuming the chelate complex Ca-ATP2- to be a substrate of the enzyme. The results obtained show that if the chelate complex formed in the aqueous stage is a substrate, then, a competition for adsorption at the active centre is possible between Ca-ATP2- and ATP4- but not between Ca-ATP2 and Ca2+. If the substrate is the chelate complex formed directly at the active centre, then ATP4- and the enzyme must combine through ion Ca2+ according to the scheme: enzyme - Ca2+ - ATP4-.

[Enzymatic properties in muscle membranes].

A study in the enzymatic properties of muscle membranes established that sarcolemma of the rabbit skeletal muscles contains the Ca2+-ATPase system which does not require Mg2+ for manifestation of ions activity. By some kinetic properties it differs from ATPase of myosin. The complex Ca-ATP2+ is a substrate of Ca2+-ATPase. Ions of a series of bivalent metals inhibit the latter as well as the passive transport of Ca2+, that may evidence for a definite relation of Ca2+-ATPase with Ca+2 transport in skeletal muscles. Acetyl cholinesterase and AMP-aminohydrolase are strongly bound with the sarcolemma. The sarcolemma structural organization is shown to play a certain role in manifestation of their activity. On the basis of the data obtained when studying the activity in the ATPase systems and dynamics of formation and decay of the intermediate phosphorylated product in the microsomal fraction of cow and rabbit myometrium certain peculiarities are established for the active mechanisms of Ca2+ transport in smooth muscles. A problem is under discussion on the possible active participation of sarcolemma in regulation of Ca2+ concentration in the smooth muscle cells. Two ATPase systems, Mg2+-dependent and Mg2+-dependent Ca2+ activated are found in nuclei; the role of lipids of the skeletal muscles in manifestation of their activity is studied. AMP-amino hydrolase properties are characterized for different areas of the sarcoplasmatic reticulum membranes. The model of E-avitaminous muscular distrophy was used to show disturbances in the structure of sarcolemma and membranes of the sarcoplasmatic reticulum which are accompanied by changes in their ATPase and Ca2+-transporting properties.