[Preparation of the myristoylated and nonmyristoylated form of recombinant recoverin in E. coli cells and comparison of their functional activity].

A recombinant plasmid was constructed for expressing a gene for bovine recoverin under the control of the lac promoter. Coexpression of the recoverin and N-myristoyl transferase genes was performed to prepare recombinant myristoylated recoverin. The obtained systems provide high levels of biosynthesis of the recombinant recoverins in the E. coli cells. Using a reconstructed system, containing urea-washed rod outer segment membranes, purified rhodopsin kinase (RK), and a recoverin, it was shown that the three recoverin forms (natural, recombinant nonmyristoylated, and recombinant myristoylated ones) perform the calcium-dependent regulation of the activity of RK with half a maximum effect at a free calcium concentration of 2 microM. Interestingly, the N-terminal myristoylation of recoverin increased substantially its functional activity.