mRNA-dependent synthesis of rat apolipoprotein E in vitro: cotranslational processing and identification of an endoglycosidase H-sensitive glycopeptide intermediate.
mRNA-dependent synthesis of rat apolipoprotein E in vitro: cotranslational processing and identification of an endoglycosidase H-sensitive glycopeptide intermediate.
复制标题
大鼠载脂蛋白 E 体外 mRNA 依赖性合成：内切糖苷酶 H 敏感糖肽中间体的共翻译加工和鉴定。
DOI：
10.1016/0006-291x(81)91794-0
复制
发表时间：
1981
影响因子：
3.1
通讯作者：
L. Chan
中科院分区：
文献类型：
--
作者：
Y. Lin;W. Bradley;L. Chan
用盐酸胍提取和oligo（dT）-纤维素层析法从大鼠肝脏中制备总聚腺苷酸化富集mRNA。它在体外翻译的mRNA依赖的小麦胚芽系统和兔网织红细胞裂解物系统，使用放射性标记的亮氨酸或蛋氨酸作为氨基酸前体。一个产品，指定preapoE，特异性沉淀的兔抗大鼠apoE血清，占总放射性肽的1.5%。在SDS凝胶上以单一放射性条带迁移，表观分子量与成熟血浆apoE相似。在翻译反应中包含犬胰腺微粒体膜导致产物略小（500道尔顿）。它还将preapoE从糖苷内切酶H抗性转化为酶敏感性物种。这表明前apoE的加工通过信号肽的共翻译去除和成熟蛋白的核心糖基化而发生。
Total polyadenylated enriched mRNA was prepared from rat liver by guanidine-HCl extraction and oligo (dT)-cellulose chromatography. It was translated in vitro in an mRNA-dependent wheat germ system and rabbit reticulocyte lysate system, using radiolabeled leucine or methionine as amino acid precursor. A product, designated preapoE, was specifically precipitated by a rabbit anti-rat apoE serum and accounted for 1.5% of the total radioactive peptides. It migrated as a single band of radioactivity on SDS gels with an apparent molecular weight similar to that of mature plasma apoE. Inclusion of dog pancreatic microsomal membranes in the translation reaction resulted in a slightly smaller product (by 500 daltons). It also converted the preapoE from an endoglycosidase H-resistant to an enzyme-sensitive species. This suggests that processing of preapoE takes place by the cotranslational removal of a signal peptide and core glycosylation of the mature protein.