Fragmentation of biotinylated cyclic peptides

被引:5
作者
Lassman, ME [1 ]
Kulagina, N [1 ]
Taitt, CR [1 ]
机构
[1] USN, Res Lab, Ctr Biomol Sci & Engn, Washington, DC 20375 USA
关键词
D O I
10.1002/rcm.1483
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Electrospray ionization coupled with tandem mass spectrometry (MS/MS) was used to determine the preferred binding site(s) of biotin NHS ester with a series of cyclic peptides with antibiotic properties. The peptides investigated are polymyxins, cyclic peptides produced by Bacillus polymyxa. In spite of the 1:1 stoichiometry used in the labeling reaction, multiple biotin molecules were incorporated into intact polymyxin peptides. Given the amine specificity of the activated biotin and the large number of amino acids with primary amines in the polymyxins, it was not clear by inspection which binding sites were more reactive than others. MS/MS was used to characterize the structure of the biotinylated peptides. MS/MS spectra of cyclic peptides often lead to ambiguous structure determinations due to the potential for multiple ring openings which result in the generation of multiple ion series. The MS/MS spectra of polymyxin peptides are especially difficult to characterize due to the lack of variety in their amino acids; however, the added complexity of the biotin aided the elucidation of the fragmentation pathways. MS/MS spectra of the species with biotin additions were used to rationalize the preferential binding sites of these molecules. Published in 2004 by John Wiley Sons, Ltd.
引用
收藏
页码:1277 / 1285
页数:9
相关论文
共 17 条
[1]   AEROSPORIN, AN ANTIBIOTIC PRODUCED BY BACILLUS-AEROSPORUS GREER [J].
AINSWORTH, GC ;
BROWN, AM ;
BROWNLEE, G .
NATURE, 1947, 160 (4060) :263-263
[2]  
BENEDICT RG, 1947, J BACTERIOL, V54, P24
[3]   NOMENCLATURE FOR PEPTIDE FRAGMENT IONS (POSITIVE-IONS) [J].
BIEMANN, K .
METHODS IN ENZYMOLOGY, 1990, 193 :886-887
[4]   PROTECTION FROM ENDOTOXEMIA - A RAT MODEL OF PLASMAPHERESIS AND SPECIFIC ADSORPTION WITH POLYMYXIN-B [J].
COHEN, J ;
ASLAM, M ;
PUSEY, CD ;
RYAN, CJ .
JOURNAL OF INFECTIOUS DISEASES, 1987, 155 (04) :690-695
[5]   Liquid chromatography-ion trap tandem mass spectrometry for the characterization of polypeptide antibiotics of the colistin series in commercial samples [J].
Govaerts, C ;
Orwa, J ;
Van Schepdael, A ;
Roets, E ;
Hoogmartens, J .
JOURNAL OF CHROMATOGRAPHY A, 2002, 976 (1-2) :65-78
[6]   Mass spectrometric fragmentation of cyclic peptides belonging to the polymyxin and colistin antibiotics studied by ion trap and quadrupole/orthogonalacceleration time-of-flight technology [J].
Govaerts, C ;
Rozenski, J ;
Orwa, J ;
Roets, E ;
Van Schepdael, A ;
Hoogmartens, J .
RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 2002, 16 (09) :823-833
[7]  
HANDLEY D, 1993, Patent No. 5177059
[8]   REMOVAL OF GRAM-NEGATIVE ENDOTOXIN FROM SOLUTIONS BY AFFINITY-CHROMATOGRAPHY [J].
ISSEKUTZ, AC .
JOURNAL OF IMMUNOLOGICAL METHODS, 1983, 61 (03) :275-281
[9]   Detection of endotoxin using an evanescent wave fiber-optic biosensor [J].
James, EA ;
Schmeltzer, K ;
Ligler, FS .
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, 1996, 60 (03) :189-202
[10]  
LIGLER FS, 2003, IN PRESS SPIE