Identification and characterization of recombinant murine interleukin-6 with a C-terminal pentapeptide extension using capillary reversed phase HPLC-MS and Edman degradation

被引：1

作者：

Hammacher, A ^{[1
]}

Reid, GE ^{[1
]}

Moritz, RL ^{[1
]}

Simpson, RJ ^{[1
]}

机构：

[1] WALTER & ELIZA HALL INST MED RES,LUDWIG INST CANC RES,JOINT PROT STRUCT LAB,PARKVILLE,VIC 3050,AUSTRALIA

来源：

BIOMEDICAL CHROMATOGRAPHY | 1997年 / 11卷 / 06期

关键词：

ESI-MS; recombinant IL-6; C-terminal mutant; capillary RP-HPLC;

D O I：

10.1002/(SICI)1099-0801(199711)11:6<337::AID-BMC687>3.3.CO;2-5

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We have identified a preparation of recombinant murine interleukin-6 (mIL-6) that, in addition to the anticipated product, also contained approximately equal amounts of mIL-6 with a C-terminal pentapeptide extension, The extension mutant was generated by readthrough of the stopcodon, and termination at a second in-frame stopcodon 12 base pairs 3' in the expression vector. Aliquots of the preparation were subjected to proteolytic digestion with Asp-N and Lys-C-endopeptidase, The resultant peptides were separated by reversed-phase capillary HPLC, and analysed using a combination of mass spectrometry and N-terminal sequence analysis, These data revealed a C-terminal pentapeptide (Gln-Gly-Ser-Val-Asp) extension, with the authentic stopcodon being translated as glutamine, The extension mutant was isolated by reversed-phase HPLC and shown to have similar mitogenic activity to mIL-6 on murine hybridoma 7TD1 cells. (C) 1997 John Wiley & Sons, Ltd.

引用

页码：337 / 342

页数：6