Characterization of calmodulin and calmodulin isotypes from sea urchin gametes.
Characterization of calmodulin and calmodulin isotypes from sea urchin gametes.
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DOI：
10.1016/s0021-9258(19)68108-1
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发表时间：
1982-02
期刊：
影响因子：
--
通讯作者：
W. Burgess
中科院分区：
文献类型：
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作者：
W. Burgess
开发了从海胆 Strongylocentrotus purpuratus 和 Arbacia punctulata 的卵和精子中纯化钙调蛋白的程序。从点状蝽卵中分离出两种形式的钙调蛋白，其他三个来源各分离出一种。所有五种钙调蛋白都与脊椎动物钙调蛋白相似，根据其对牛脑环核苷酸磷酸二酯酶的激活、与肌钙蛋白 I 和氯丙嗪的钙依赖性相互作用、钙存在下十二烷基硫酸钠凝胶上的阳极迁移率增加、与抗脊椎动物钙调蛋白抗体和氨基酸组合物（包括存在三甲基赖氨酸的单个残基）的交叉反应性来判断。斑点毛蠓卵中的两种形式的钙调蛋白，即钙调蛋白 A 和钙调蛋白 B，可以根据 DEAE-纤维素色谱法、氨基酸分析、有或无钙的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳以及竞争性放射免疫测定法来区分。没有证据表明任何一种形式是另一种形式的蛋白水解产物。
Procedures were developed for the purification of calmodulin from the eggs and from the sperm of the sea urchin Strongylocentrotus purpuratus and Arbacia punctulata. Two forms of calmodulin were isolated from A. punctulata eggs, one from each of the other three sources. All five calmodulins are similar to vertebrate calmodulin as judged by their activation of bovine brain cyclic nucleotide phosphodiesterase, calcium-dependent interaction with troponin I and with chloropromazine, increased anode mobility on sodium dodecyl sulfate gels in the presence of calcium, cross-reactivity with anti-vertebrate calmodulin antibodies and amino acid compositions including the presence of a single residue of trymethyllysine. The two forms of calmodulin from A. punctulata eggs, calmodulin A and calmodulin B, can be distinguished from one another on the basis of DEAE-cellulose chromatography, amino acid analysis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence or absence of calcium and competitive radioimmunoassays. There is no evidence that either form is a proteolytic product of the other.