Actin structure-function relationships in vitro using oligodeoxynucleotide-directed site-specific mutagenesis.
Actin structure-function relationships in vitro using oligodeoxynucleotide-directed site-specific mutagenesis.
复制标题
使用寡脱氧核苷酸定向位点特异性诱变在体外研究肌动蛋白结构-功能关系。
DOI：
10.1002/cm.970140108
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发表时间：
1989
影响因子：
--
通讯作者：
Gay,L
中科院分区：
文献类型：
--
作者：
Rubenstein,PA;Solomon,LR;Solomon,T;Gay,L
肌动蛋白是每个真核细胞中发现的主要结构和收缩蛋白。肌动蛋白的晶体结构尚未完全阐明；然而，已经报道了肌动蛋白：DNAse I 复合物的 4.5 A 图谱，该图显示肌动蛋白似乎由大的羧基末端结构域和小的氨基末端结构域组成[Kabsch et al., 19851。其氨基酸结构在整个进化过程中保持高度保守。这种情况可能允许在活细胞和无细胞系统中用一种肌动蛋白替代另一种肌动蛋白，以便研究肌动蛋白功能的不同方面。肌动蛋白通常以两种方式进行翻译后修饰。首先，迄今为止研究的每个肌动蛋白的第 73 位都有一个不寻常的残基，即 3-甲基组氨酸 [Collins 和 Elzinga, 19751，但来自 Nuegleria gruberi 的肌动蛋白除外，其中该位置的组氨酸保持未甲基化 [Sussman, 19841]。其次，在迄今为止研究的所有肌动蛋白中，可能除了来自 Acun-thamoebacastellani 的肌动蛋白 [Redman 等人， 19851，肌动蛋白被合成为具有一到四个额外 NH2 末端氨基酸的前体分子，其中大部分似乎以乙酰化依赖性方式被去除[Redman 和 Rubenstein，1981；鲁宾斯坦和马丁，1983a，b； Solomon 和 Rubenstein，19851。这些修饰的生物学意义仍然未知。细胞内的肌动蛋白功能部分通过其与大量细胞蛋白的相互作用进行调节[Pollard 和 Cooper，19861。为了确定肌动蛋白与各种肌动蛋白结合蛋白相互作用的方式，已经进行了利用零长度交联剂的实验，试图识别这些蛋白在肌动蛋白表面上的结合位点。这些研究表明，在存在 EDAC (1-(3-dime-
Actin is a major structural and contractile protein found in every eukaryotic cell. The crystal structure of actin has not been completely elucidated; however, a 4.5 A map of the actin: DNAse I complex, which shows that the actin appears to be composed of large carboxyl-terminal domain and a small amino-terminal domain, has been reported [Kabsch et al., 19851. Its amino acid structure has remained highly conserved throughout evolution. This situation potentially allows for the substitution of one actin for another in both living and cell-free systems in order to study different aspects of actin function. Actin is usually post-translationally modified in two ways. First, there is an unusual residue, 3-methylhistidine, at position 73 of every actin studied thus far [Collins and Elzinga, 19751, with the exception of that from Nuegleria gruberi in which the histidine at that position remains unmethylated [Sussman, 19841. Second, in all actins studied to date, except perhaps that from Acun-thamoeba castellani [Redman et al., 19851, the actin is synthesized as a precursor molecule with one to four extra NH2-terminal amino acids, most of which appear to be removed in an acetylation-dependent fashion [Redman and Rubenstein, 1981; Rubenstein and Martin, 1983a, b; Solomon and Rubenstein, 19851. The biological significance of these modifications remains unknown.Actin function within the cell is regulated in part through its interactions with a large number of cellular proteins [Pollard and Cooper, 19861. To determine the ways in which actin interacts with various actin-binding proteins, experiments utilizing zero-length cross-linkers have been carried out to try to identify binding sites for these proteins on the actin surface. These studies have demonstrated that in the presence of EDAC (1-(3-dime-