Regulation of Functional Activity of Adenylyl Cyclase Signal System by Synthetic Coil-Forming Peptides in Mouse Fibroblast Culture of Line L (ISM Subline)

Abstract

The signal transduction process via adenylyl cyclase system (ACS) requires coordinated functioning of signal proteins—components of ACS. It is suggested that functional coupling between them, together with other molecular mechanisms, is based on coiled-coil interactions. To study role of these interactions in functioning of ACS, we synthesized cationic coiled-forming peptides with a regular structure Ac–Ala–His– (Ala)2–His–Ala–NH2 (I), Ac–Ala–His–(Ala)3–His– (Ala)2–His–Ala–NH2 (II), and Ac–(Pro(2–His– (Ala)2–His– (Ala)2–His– (Ala)2–His–Ala–NH2 (III). Using circular dichroism (CD) spectroscopy, a portion of α-helix conformation in their secondary structure was determined, and effects of these peptides on basal adenylyl cyclase (AC) activity as well as on the activity stimulated by non-hormonal (NaF and Gpp[NH]p) and hormonal (serotonin) agents was studied in homogenate of mouse fibroblasts, line L (subline LSM). The synthetic peptides were shown to inhibit in a dose-dependent manner both basal and induced AC activity, which indicates their uncoupling action on ACS. The biological effect of these peptides correlated with their length (I < II < III), but not with coiled-coil structure, which was 20, 7, and 21%, respectively, according to data of circular dichroism spectroscopy in 3-fluoroethanol. However, there are reasons to believe that the coiled-coil structure of peptides, first place extended ones, increases at interaction with plasma membrane and signal proteins, which affects the degree of their effect on functional ACS activity. At micromolar concentrations, peptides II and III were established to markedly stimulate the basal AC activity, thereby mimicking G-protein-binding sites of cytoplasmic receptor loops. The data obtained indicate participation of the coiled-coil interactions in functional coupling of ACS components, and the methodology itself of the use of model peptides with different coiled-coil structure and distribution of charged amino acids is an efficient approach for studying molecular bases for functioning of signal systems.