Peptide-mass profiles of polyvinylidene difluoride-bound proteins by matrix-assisted laser desorption ionization time-of-flight mass spectrometry in the presence of nonionic detergents

被引:92
作者
Gharahdaghi, F
Kirchner, M
Fernandez, J
Mische, SM
机构
[1] Rockefeller University Protein, DNA Technology Center, Box 105, New York, NY 10021
基金
美国国家卫生研究院;
关键词
D O I
10.1006/abio.1996.0012
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), in conjunction with enzymatic digestion of proteins and molecular weight search of peptide-mass database is a powerful technique for peptide/protein identification. Ideally, peptide mixtures should be compatible with both MALDI-TOF and microsequencing. In our laboratory, enzymatic digestion and extraction of peptides from polyvinylidene difluoride (PVDF)bound proteins is performed in the presence of nonionic detergents. However, nonionic detergents have been shown to cause signal suppression in MALDI-TOF analysis. This study demonstrates that by using a modified matrix solution, peptide-mass fingerprinting of PVDF-bound proteins by MALDI-TOF can be obtained in the presence of nonionic detergents such as hydrogenated Triton X-100 (RTX-100), octylglucopyranoside, and Tween 20. (C) 1996 Academic Press, Inc.
引用
收藏
页码:94 / 99
页数:6
相关论文
共 28 条
[1]   ALPHA-CYANO-4-HYDROXYCINNAMIC ACID AS A MATRIX FOR MATRIX-ASSISTED LASER DESORPTION MASS-SPECTROMETRY [J].
BEAVIS, RC ;
CHAUDHARY, T ;
CHAIT, BT .
ORGANIC MASS SPECTROMETRY, 1992, 27 (02) :156-158
[2]  
BEST S, 1994, TECHNIQUES PROTEIN C, V5, P205
[3]   ALKYLATION OF CYSTEINE WITH ACRYLAMIDE FOR PROTEIN-SEQUENCE ANALYSIS [J].
BRUNE, DC .
ANALYTICAL BIOCHEMISTRY, 1992, 207 (02) :285-290
[4]   HIGH-SENSITIVITY SEQUENCING OF LARGE PROTEINS - PARTIAL STRUCTURE OF THE RAPAMYCIN-FKBP12 TARGET [J].
ERDJUMENTBROMAGE, H ;
LUI, M ;
SABATINI, DM ;
SNYDER, SH ;
TEMPST, P .
PROTEIN SCIENCE, 1994, 3 (12) :2435-2446
[5]  
FERNANDEZ J, 1995, TECH PROT CHEM, V6, P135, DOI 10.1016/S1080-8914(06)80019-0
[6]   AN IMPROVED PROCEDURE FOR ENZYMATIC DIGESTION OF POLYVINYLIDENE DIFLUORIDE-BOUND PROTEINS FOR INTERNAL SEQUENCE-ANALYSIS [J].
FERNANDEZ, J ;
ANDREWS, L ;
MISCHE, SM .
ANALYTICAL BIOCHEMISTRY, 1994, 218 (01) :112-117
[7]   INTERNAL PROTEIN-SEQUENCE ANALYSIS - ENZYMATIC DIGESTION FOR LESS THAN 10 MU-G OF PROTEIN-BOUND TO POLYVINYLIDENE DIFLUORIDE OR NITROCELLULOSE MEMBRANES [J].
FERNANDEZ, J ;
DEMOTT, M ;
ATHERTON, D ;
MISCHE, SM .
ANALYTICAL BIOCHEMISTRY, 1992, 201 (02) :255-264
[8]  
FERNANDEZ J, 1994, TECHNIQUES PROTEIN C, V6, P215
[9]  
GHARAHDAGHI F, 1995, 43RD P ASMS C MASS S, P918
[10]   IDENTIFYING PROTEINS FROM 2-DIMENSIONAL GELS BY MOLECULAR MASS SEARCHING OF PEPTIDE-FRAGMENTS IN PROTEIN-SEQUENCE DATABASES [J].
HENZEL, WJ ;
BILLECI, TM ;
STULTS, JT ;
WONG, SC ;
GRIMLEY, C ;
WATANABE, C .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (11) :5011-5015