Purification and kinetic properties of two soluble forms of calmodulin-dependent cyclic nucleotide phosphodiesterase from rat pancreas.

The calmodulin-dependent cyclic AMP phosphodiesterase and cyclic GMP phosphodiesterase (EC 3.1.4.17) activity of rat pancreas was purified 280-fold by affinity chromatography on calmodulin-Sepharose 4B. It then accounted for 15% of the total cytosol cyclic GMP nucleotide phosphodiesterase activity, in the presence of Ca2+, and represented a minor component of proteins specifically adsorbed by the column. This activity was resolved on a DEAE-Sephacel column into two fractions, termed PI and PII, on the basis of their order of emergence. After this step, PI and PII were purified 5650- and 3700-fold respectively. The molecular weight of PI was 175 000 and that of PII was 116 000, by polyacrylamide-gradient-gel electrophoresis. Both forms of phosphodiesterase could hydrolyse cyclic AMP and cyclic GMP, although PII displayed a higher affinity toward cyclic GMP than toward cyclic AMP. PI and PII exhibited negative homotropic kinetics in the absence of calmodulin. Upon addition of calmodulin, both enzymes displayed Michaelis-Menten kinetics and a 5-9-fold increase in maximal velocity, at physiological concentrations of cyclic GMP and cyclic AMP. When a pancreatic extract freshly purified by affinity chromatography was immediately analysed by high-performance gel-permeation chromatography on a TSK gel G3000 SW column, PII represented as much as 78% of the eluted activity. This percentage decreased to 52% when the sample was stored at 0 degrees C for 20 h before analysis, suggesting that PII, possibly predominant in vivo, was converted into the heavier PI form upon storage

Chemical, immunological and biological properties of peptides like vasoactive-intestinal-peptide and peptide-histidine-isoleucinamide extracted from the venom of two lizards (Helodermahorridum and Helodermasuspectum).

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Inhibition of Na/Ca exchange by Phe-Met-Arg-Phe-NH2 (FMRFa)-related peptides in intact rat pancreatic B-cells.

Phe-Met-Arg-Phe-NH2 (FMRFa)-related peptides were recently shown to inhibit Na/Ca exchange in cardiac sarcolemmal vesicles. In the present study, we examined the effects of FMRFa-related peptides on Na/Ca exchange in intact (pancreatic B) cells. At 2.8 mM glucose, FMRFa-related peptides only weakly inhibited Na/Ca exchange although their effect was more marked under depolarizing conditions. The peptides blocked neither the Na/K-ATPase nor Ca2+ channels but slightly reduced membrane K+ permeability. Our data indicate that FMRFa-related peptides are weak and non-specific inhibitors of Na/Ca exchange in intact B cells. The data do not confirm the view that the peptides may exert some of their physiological modulatory role by inhibiting Na/Ca exchange.Journal ArticleResearch Support, Non-U.S. Gov'tinfo:eu-repo/semantics/publishe

Effects of Phe-Met-Arg-Phe-NH2 (FMRFa)-related peptides on Na-Ca exchange and ionic fluxes in rat pancreatic B cells

Journal ArticleFLWNAinfo:eu-repo/semantics/publishedSodium–Calcium Exchange: Proceedings of the Third International Conferenc

Phe-Met-Arg-Phe-NH2 (FMRFa)-related peptides inhibit Na/Ca exchange in pancreatic B cells.

Journal Articleinfo:eu-repo/semantics/publishe