Terashima M, Yamamori C, Shimoyama M
Department of Biochemistry, Shimane Medical University, Izumo, Japan.
Eur J Biochem. 1995 Jul 1;231(1):242-9.
We reported previously on ADP-ribosylation of actins by chicken arginine-specific ADP-ribosyltransferase in vitro and in situ and the inhibition of actin polymerization by this modification [Terashima, M., Mishima, K., Yamada, K., Tsuchiya, M., Wakutani, T. & Shimoyama, M. (1992) Eur. J. Biochem. 204, 305-311]. In the present study, we determined amino acid residues of ADP-ribosylation site(s) in globular (G-) and filamentous (F-) actins and examined the molecular basis of the modification of actin. Arginine-specific ADP-ribosylation occurred at Arg28 and Arg206 in G-actin, but only at Arg28 in F-actin. ADP-ribosylation of Arg206, located on the pointed end of the actin molecule, significantly blocked the interaction with deoxyribonuclease I. These results indicate that Arg206 in G-actin may be involved in actin polymerization. ADP-ribosylation of Arg28, located on the outer surface of actin molecule, did not affect the binding activity with myosin subfragment-1, that is thought to interact through the N-terminal amino acid residues of G-actin. ADP-ribosylation at both Arg28 and Arg206 of G-actin had no apparent effect on the intrinsic ATPase activity. We concluded from this study that ADP-ribosylation of Arg206 in G-actin causes the inhibition of actin polymerization, and that ADP-ribosylation of Arg28 occurs in F-actin.
我们之前报道过鸡精氨酸特异性 ADP - 核糖基转移酶在体外和原位对肌动蛋白进行 ADP - 核糖基化作用，以及这种修饰对肌动蛋白聚合的抑制作用[寺岛，M.，三岛，K.，山田，K.，土屋，M.，若谷，T. & 下山，M.（1992 年）《欧洲生物化学杂志》204 卷，305 - 311 页]。在本研究中，我们确定了球状（G -）和丝状（F -）肌动蛋白中 ADP - 核糖基化位点的氨基酸残基，并研究了肌动蛋白修饰的分子基础。精氨酸特异性 ADP - 核糖基化发生在 G - 肌动蛋白的 Arg28 和 Arg206 位点，但在 F - 肌动蛋白中仅发生在 Arg28 位点。位于肌动蛋白分子尖端的 Arg206 的 ADP - 核糖基化显著阻断了与脱氧核糖核酸酶 I 的相互作用。这些结果表明 G - 肌动蛋白中的 Arg206 可能参与肌动蛋白聚合。位于肌动蛋白分子外表面的 Arg28 的 ADP - 核糖基化不影响与肌球蛋白亚片段 - 1 的结合活性，据认为肌球蛋白亚片段 - 1 通过 G - 肌动蛋白的 N 端氨基酸残基相互作用。G - 肌动蛋白的 Arg28 和 Arg206 位点的 ADP - 核糖基化对内在 ATP 酶活性没有明显影响。我们从这项研究得出结论，G - 肌动蛋白中 Arg206 的 ADP - 核糖基化导致肌动蛋白聚合的抑制，并且 Arg28 的 ADP - 核糖基化发生在 F - 肌动蛋白中。
