人二磷酸甘油酸变位酶信使核糖核酸的无细胞翻译

Cell-free translation of messenger RNA for human bisphosphoglyceromutase.

作者信息

Dubart A, Romeo P H, Tsapis A, Goossens M, Rosa R, Rosa J

出版信息

Biochem Biophys Res Commun. 1984 Apr 30;120(2):441-7. doi: 10.1016/0006-291x(84)91273-7.

Abstract

mRNAs prepared from different human tissues were translated in a cell-free reticulocyte lysate system and, when present, the neosynthesized bisphosphoglyceromutase ( BPGM ) was specifically isolated by immunoprecipitation and analyzed by polyacrylamide gel electrophoresis. Analysis of the translation products showed that bisphosphoglyceromutase was synthesized in vitro with its mature molecular weight and messenger RNA specifying the synthesis of BPGM exhibited a sedimentation coefficient of 12 S in human reticulocytes. This synthesis seems to be highly tissue specific since we could not evidence any synthesis of this enzyme using mRNA obtained from non erythroid tissue. The BPGM synthesis represents 0.1% of the total neosynthesized non heme proteins in human reticulocyte and ten times less (0.01%) in human fetal liver.

摘要

从不同人体组织制备的mRNA在无细胞网织红细胞裂解物系统中进行翻译，如有新合成的二磷酸甘油酸变位酶（BPGM），则通过免疫沉淀法进行特异性分离，并通过聚丙烯酰胺凝胶电泳进行分析。对翻译产物的分析表明，二磷酸甘油酸变位酶在体外以其成熟分子量合成，且在人网织红细胞中，指定BPGM合成的信使RNA沉降系数为12S。这种合成似乎具有高度的组织特异性，因为我们使用从非红细胞组织获得的mRNA未能证明该酶的任何合成。BPGM的合成占人网织红细胞中新合成的非血红素蛋白总量的0.1%，而在人胎儿肝脏中则少十倍（0.01%）。