S100B（ββ）与源自p53的肽之间的钙依赖性相互作用。

The Ca(2+)-dependent interaction of S100B(beta beta) with a peptide derived from p53.

作者信息

Rustandi R R, Drohat A C, Baldisseri D M, Wilder P T, Weber D J

机构信息

Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore 21201, USA.

出版信息

Biochemistry. 1998 Feb 17;37(7):1951-60. doi: 10.1021/bi972701n.

DOI:10.1021/bi972701n

PMID:9485322

Abstract

S100B(beta beta) was found to interact with the tumor suppressor protein, p53, and inhibit its PKC-dependent phosphorylation and tetramer formation [Baudier, J., Delphin, C., Grunwald, D., Khochbin, S., and Lawrence, J. J. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 11627-11631]. Since PKC-dependent phosphorylation at the C-terminus of p53 is known to effect transcription and p53 tetramer formation [Sakaguchi, K., Sakamoto, H., Lewis, M. S., Anderson, C. W., Erickson, J. W., Appella, E., and Xie, D. (1997) Biochemistry 36, 10117-10124], we examined the interaction of S100B(beta beta) with a peptide derived from the C-terminal regulatory domain of p53 (residues 367-388). In this paper, we report that S100B(beta beta) binds to the p53 peptide (CaK3 < or = 23.5 +/- 6.6 microM) in a Ca(2+)-dependent manner, and that the presence of the p53 peptide was found to increase the binding affinity of Ca2+ to S100B(beta beta) by 3-fold using EPR and PRR methods, whereas the peptide had no effect on Zn2+ binding to S100B(beta beta). Fluorescence and NMR spectroscopy experiments show that the p53 peptide binds to a region of S100B(beta beta) that probably includes residues in the "hinge" (S41, L44, E45, E46, I47), C-terminal loop (A83, C84, H85, E86, F87, F88), and helix 3 (V52, V53, V56, T59). Together these data support the notion that S100B(beta beta) inhibits PKC-dependent phosphorylation by binding directly to the C-terminus of p53.

摘要

研究发现，S100B（ββ）可与肿瘤抑制蛋白p53相互作用，并抑制其依赖蛋白激酶C（PKC）的磷酸化及四聚体形成[鲍迪尔，J.，德尔芬，C.，格伦瓦尔德，D.，霍奇宾，S.，以及劳伦斯，J. J.（1992年）《美国国家科学院院刊》89，11627 - 11631]。由于已知p53 C端依赖PKC的磷酸化会影响转录及p53四聚体形成[坂口，K.，坂本，H.，刘易斯，M. S.，安德森，C. W.，埃里克森，J. W.，阿佩拉，E.，以及谢，D.（1997年）《生物化学》36，10117 - 10124]，我们研究了S100B（ββ）与源自p53 C端调节结构域的肽段（第367 - 388位氨基酸残基）之间的相互作用。在本文中，我们报告S100B（ββ）以Ca（2 +）依赖的方式与p53肽段结合（CaK3≤23.5±6.6微摩尔），并且通过电子顺磁共振（EPR）和脉冲射频弛豫（PRR）方法发现，p53肽段的存在使Ca2 +与S100B（ββ）的结合亲和力提高了3倍，而该肽段对Zn2 +与S100B（ββ）的结合没有影响。荧光和核磁共振光谱实验表明，p53肽段与S100B（ββ）的一个区域结合，该区域可能包括“铰链区”（S41、L44、E45、E46、I47）、C端环（A83、C84、H85、E86、F87、F88）以及螺旋3（V52、V53、V56、T59）中的氨基酸残基。这些数据共同支持了S100B（ββ）通过直接结合p53的C端来抑制PKC依赖的磷酸化这一观点。