CHARACTERIZATION OF DISULFIDE LINKAGES AND DISULFIDE BOND SCRAMBLING IN RECOMBINANT HUMAN MACROPHAGE-COLONY-STIMULATING FACTOR BY FAST-ATOM-BOMBARDMENT MASS-SPECTROMETRY OF ENZYMATIC DIGESTS

被引：29

作者：

GLOCKER, MO

ARBOGAST, B

DEINZER, ML

机构：

[1] OREGON STATE UNIV,DEPT AGR CHEM,CORVALLIS,OR 97331

[2] OREGON STATE UNIV,CTR ENVIRONM HLTH SCI,CORVALLIS,OR 97331

[3] UNIV KONSTANZ,FAC CHEM,D-78434 CONSTANCE,GERMANY

来源：

JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY | 1995年 / 6卷 / 08期

关键词：

D O I：

10.1016/1044-0305(95)00250-H

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Fast-atom bombardment mass spectrometry was used to study disulfide bonding patterns in heat-denatured human recombinant macrophage colony stimulating factor (rhM-CSF). The heat-denaturated protein was studied by analysis of the pattern of peptides in the proteolytic digests. Native rhM-CSF is a homodimer with intramolecular disulfide linkages between Cys7-Cys90, Cys48-Cys139, and Cys102-Cys146 and intermolecular linkages between Cys31-Cys31, and the pairs Cys157 and Cys159. Brief heating for 1 min leads to partial disulfide bond scrambling. In addition to the native disulfide bonds between Cys7-Cys90, Cys48-Cys139, and Cys31-Cys31, nonnative disulfide bonds were detected between Cys48-Cys90 and Cys48-Cys102. When heated for 5 min the disulfide bonds of rhM-CSF are completely scrambled and lead to nonnative intramolecular disulfide bonds between Cys48-Cys102 and Cys90-Cys102 and one intermolecular disulfide bond between Cys102-Cys102.

引用

页码：638 / 643

页数：6

共 23 条

[1] SEQUENTIAL RESONANCE ASSIGNMENTS IN PROTEIN H-1 NUCLEAR MAGNETIC-RESONANCE SPECTRA - COMPUTATION OF STERICALLY ALLOWED PROTON PROTON DISTANCES AND STATISTICAL-ANALYSIS OF PROTON PROTON DISTANCES IN SINGLE-CRYSTAL PROTEIN CONFORMATIONS [J].