Two-dimensional analysis of recombinant E-coli proteins using capillary isoelectric focusing electrospray ionization mass spectrometry

被引:71
作者
Tang, Q
Harrata, AK
Lee, CS
机构
[1] US DOE,DEPT CHEM,AMES,IA 50011
[2] US DOE,AMES LAB,AMES,IA 50011
关键词
D O I
10.1021/ac970015o
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
On-line combination of capillary isoelectric focusing with electrospray ionization mass spectrometry is applied for a two-dimensional analysis of Escherichia coli proteins. The proteins are focused and cathodically mobilized in a polyacrylamide coated capillary. At the end of the capillary, various protein zones are analyzed by mass spectrometry coupled through an electrospray interface, Comparisons with silver-stained two-dimensional gel electrophoresis are made with regard to mass determination, resolution, speed, and sensitivity, Direct identification of a recombinant fusion protein of glutathione S-transferase and striped bass growth hormone is achieved without any prior protein isolation procedures.
引用
收藏
页码:3177 / 3182
页数:6
相关论文
共 21 条
[1]   PRODUCTION OF A BIOLOGICALLY-ACTIVE RECOMBINANT TELEOSTEAN GROWTH-HORMONE IN E-COLI CELLS [J].
CHENG, CM ;
LIN, CM ;
SHAMBLOTT, M ;
GONZALEZVILLASENOR, LI ;
POWERS, DA ;
WOODS, C ;
CHEN, TT .
MOLECULAR AND CELLULAR ENDOCRINOLOGY, 1995, 108 (1-2) :75-85
[2]  
CREIGHTON TE, 1990, PROTEIN STRUCTURE PR, pCH3
[3]   ELECTROSPRAY IONIZATION-PRINCIPLES AND PRACTICE [J].
FENN, JB ;
MANN, M ;
MENG, CK ;
WONG, SF ;
WHITEHOUSE, CM .
MASS SPECTROMETRY REVIEWS, 1990, 9 (01) :37-70
[4]   ADAPTATION OF THE EQUIPMENT FOR HIGH-PERFORMANCE ELECTROPHORESIS TO ISOELECTRIC-FOCUSING [J].
HJERTEN, S ;
ZHU, MD .
JOURNAL OF CHROMATOGRAPHY, 1985, 346 (OCT) :265-270
[5]   THEORETICAL AND EXPERIMENTAL-STUDY OF HIGH-PERFORMANCE ELECTROPHORETIC MOBILIZATION OF ISOELECTRICALLY FOCUSED PROTEIN ZONES [J].
HJERTEN, S ;
LIAO, JL ;
YAO, K .
JOURNAL OF CHROMATOGRAPHY, 1987, 387 :127-138
[6]   ELECTROSPRAY ION SPRAY - A COMPARISON OF MECHANISMS AND PERFORMANCE [J].
IKONOMOU, MG ;
BLADES, AT ;
KEBARLE, P .
ANALYTICAL CHEMISTRY, 1991, 63 (18) :1989-1998
[7]   FROM IONS IN SOLUTION TO IONS IN THE GAS-PHASE - THE MECHANISM OF ELECTROSPRAY MASS-SPECTROMETRY [J].
KEBARLE, P ;
TANG, L .
ANALYTICAL CHEMISTRY, 1993, 65 (22) :A972-A986
[8]   FAST AND HIGH-RESOLUTION ANALYSIS OF HUMAN-SERUM TRANSFERRIN BY HIGH-PERFORMANCE ISOELECTRIC-FOCUSING IN CAPILLARIES [J].
KILAR, F ;
HJERTEN, S .
ELECTROPHORESIS, 1989, 10 (01) :23-29
[9]   EFFECTS OF ANIONS ON THE POSITIVE-ION ELECTROSPRAY-IONIZATION MASS-SPECTRA OF PEPTIDES AND PROTEINS [J].
MIRZA, UA ;
CHAIT, BT .
ANALYTICAL CHEMISTRY, 1994, 66 (18) :2898-2904
[10]  
MOSHER RA, 1992, DYNAMICS ELECTROPHOR, pCH7