来自人黑素瘤细胞的佛波酯诱导的48000道尔顿蛋白的信使核糖核酸。

Messenger RNA for a phorbol-ester induced 48,000 dalton protein from human melanoma cells.

作者信息

Opdenakker G, Bosman F, Van Damme J, Billiau A

出版信息

Biochem Biophys Res Commun. 1986 Apr 14;136(1):122-9. doi: 10.1016/0006-291x(86)90885-5.

Abstract

Messenger RNA of the phorbol ester-induced 48kDa protein from human melanoma cells (Bowes) was isolated, characterized and used to study the protein processing. The 48kDa mRNA is induced simultaneously with that of tissue-type plasminogen activator. This induction is prominent as shown by sedimentation profiles on linear sucrose gradients. The mRNA can be isolated by classical phenol extractions, has a poly(A)-tail and sediments with a coefficient of 20 S. Translation in reticulocyte lysates yields a 48kDa protein whether the translation is modified with canine pancreas microsomal membranes or not. Analysis of 48kDa mRNA translation products by sodium dodecyl sulphate/polyacrylamide gel electrophoresis showed that the phorbol ester-induced 48kDa is a monomeric one-chain polypeptide. Glycosylation could not be detected, nor signal peptide cleaving, suggesting that it is a non-secreted intracellular protein.

摘要

从人黑色素瘤细胞（鲍伊斯细胞）中分离出佛波酯诱导的48kDa蛋白的信使核糖核酸（mRNA），对其进行了表征，并用于研究蛋白质加工过程。48kDa mRNA与组织型纤溶酶原激活剂的mRNA同时被诱导。如线性蔗糖梯度沉降图谱所示，这种诱导作用很显著。该mRNA可通过经典的苯酚抽提法分离得到，具有聚腺苷酸尾，沉降系数为20S。无论在网织红细胞裂解物中的翻译是否用犬胰腺微粒体膜进行修饰，都能产生一种48kDa的蛋白。通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳对48kDa mRNA翻译产物进行分析表明，佛波酯诱导的48kDa蛋白是一种单体单链多肽。未检测到糖基化现象，也未检测到信号肽裂解，这表明它是一种非分泌型细胞内蛋白。