Functional analysis of individual brain myosin II isoforms through hybrid formation.
Functional analysis of individual brain myosin II isoforms through hybrid formation.
复制标题
通过混合形成对个体脑肌球蛋白 II 亚型进行功能分析。
DOI：
10.1016/0014-5793(94)00613-x
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发表时间：
1994
期刊：
影响因子：
3.5
通讯作者：
Chantler,PD
中科院分区：
文献类型：
--
作者：
Wang,Y;Chantler,PD
我们使用了扇贝杂交肌球蛋白测试系统，试图确定大鼠脑中单个肌球蛋白II亚型的调节特性。与先前在脑中表达的调节轻链亚型相对应的完整编码区[Feinstein，Durand和Milner(1991)Mol.Brain Res.10，97-105]连接到在细菌中高效表达的原核表达载体pAED4中，并将纯化的轻链整合到扇贝杂交肌球蛋白中。无论轻链磷酸化是在杂交体形成之前还是之后发生的，肌动蛋白的激活对所有被测试的杂交体都是不敏感的。我们讨论了这些结果的含义，包括这些结果可能构成了大脑中肌球蛋白II亚型在细丝水平上受到调节的证据。此外，有证据表明，在大鼠大脑中表达了一种额外的、新的调节轻链亚型。
We have used a scallop hybrid myosin test system in an attempt to determine the regulatory properties of an individual myosin II isoform from rat brain. The complete coding region of cDNA corresponding to a regulatory light chain isoform previously shown to be expressed in brain [Feinstein, Durand and Milner (1991) Mol. Brain Res. 10, 97‐105] was ligated within the prokaryotic expression vector,pAED4, overexpressed in bacteria, and the purified light chain incorporated within a scallop hybrid myosin. Actin activation was calcium insensitive for all hybrids tested, irrespective of whether light chain phosphorylation had taken place before, or subsequent to, hybrid formation. We discuss the implications of these results, including the possibility that these results constitute evidence for a myosin II isoform within brain that is regulated at the level of the thin filament. In addition, evidence is presented for the presence of an additional, novel isoform of regulatory light chain expressed in rat brain.