In vitro biosynthesis of the beta-subunit of the Na+/K+-ATPase in developing brine shrimp: glycosylation and membrane insertion.
In vitro biosynthesis of the beta-subunit of the Na+/K+-ATPase in developing brine shrimp: glycosylation and membrane insertion.
复制标题
发育丰年虾中 Na /K -ATP 酶 β 亚基的体外生物合成：糖基化和膜插入。
DOI：
10.1016/0005-2736(88)90566-4
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Hokin,LE
中科院分区：
文献类型：
--
作者：
Baxter-Lowe,LA;Yohanan,JM;Hokin,LE
我们在这里展示了发育中的卤虫在网织红细胞裂解物翻译系统中的Na+/K+-β酶亚基的翻译、糖基化和膜插入。经SDS-PAGE测定，初步翻译产物的表观分子量为33000±1000(n=7)。当微体膜在整个转化期内存在时，出现一条表观分子量为37000±1000(n=7)的新条带。这种表观分子量的变化是由于添加了大约两个N-连接的寡糖。还研究了蛋白质合成和糖基化之间的时间关系。如果在完成约70%的多肽链后加入膜，则可以实现糖基化和膜插入。然而，如果在β亚基翻译完成后添加膜，则不会发生糖基化。β-亚基是在膜结合的多聚体上合成的，在生长的多肽链上增加了大约两个N-连接的寡糖。这些研究表明，体外翻译系统将有助于研究β-亚基的生物合成，这是研究Na+/K+-ATPase发育规律的一个很好的模型系统。
We demonstrate here translation, glycosylation, and membrane insertion of the β-subunit of the Na+/K+-ATPase of the developing brine shrimp,Artemia, in a reticulocyte lysate translation system. The apparent molecular weight of the primary translation product as determined by SDS-PAGE is 33 000 ± 1000 (n= 7). When microsomal membranes are present during the entire translation period, a new band with an apparent molecular weight of 37 000 ± 1000 (n= 7) appears. This change in apparent molecular weight is due to the addition of about two N-linked oligosaccharides. The temporal relationship between protein synthesis and glycosylation have also been examined. Glycosylation and membrane insertion could be achieved if membranes were added after completion of about 70% of the peptide chain. However, glycosylation did not occur if membranes were added after the completion of translation of the β-subunit. The β-subunit was synthesized on membrane-bound polysomes, where about two N-linked oligosaccharides were added to the growing polypeptide chain. These studies demonstrate that in vitro translation systems will be useful for studying the biosynthesis of the β-subunit of the brine shrimp, which is a good model system to examine the developmental regulation of the Na+/K+-ATPase.