ADP-RIBOSYLTRANSFERASE TYPE-A FROM TURKEY ERYTHROCYTES MODIFIES ACTINAT ARG-95 AND ARG-372

Turkey erythrocyte ADP-ribosyltransferase A catalyzes the transfer of ADP-ribose from NAD to both monomeric and polymeric skeletal muscle al pha-actin with the incorporation of 2 mol of ADP-ribose per mol of act in. In contrast, Clostridium perfringens iota toxin ADP-ribosylates on ly G-actin, with modification at arginine-177 [Vandekerckhove, J., et al. (1987) FEES Lett. 255, 48-42]. Transferase A-catalyzed modificatio ns are sensitive to 0.5 M neutral hydroxylamine, consistent with the a rginine side chain modification. Radiolabeled peptides ADP-ribosylated by transferase A were generated by tryptic digestion and purified by reversed phase high performance Liquid chromatography. Amino acid sequ ence and molecular mass analysis identified the ADP-ribosylation sites as Arg-95 and Arg-372 of actin; both residues are located within subd omain-1 of the actin 3D structure [Kabsch, W., et al. (1990) Nature 34 7, 37-44]. ADP-ribosylation did not affect cytochalasin D-stimulated G -actin ATPase, the binding of actin to DNase I or to gelsolin, or the ability of actin to polymerize. Following ADP-ribosylation, however, a prolonged delay in polymerization was observed, consistent with a dec reased rate of nucleation.