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Analysis of phospholipase A 2 glycosylation patterns from venom of individual bees by capillary electrophoresis/electrospray ionization mass spectrometry using an ion trap mass spectrometer
Author(s) -
Lai ChienChen,
Her GuorRong
Publication year - 2000
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/1097-0231(20001115)14:21<2012::aid-rcm126>3.0.co;2-8
Subject(s) - chemistry , chromatography , mass spectrometry , electrospray ionization , capillary electrophoresis , sample preparation in mass spectrometry , capillary electrophoresis–mass spectrometry , protein mass spectrometry , electrospray , tandem mass spectrometry , trypsin , biochemistry , enzyme
A method based on tryptic digestion, ultrafiltration and capillary electrophoresis/mass spectrometry (CE/MS) has been developed for the analysis of the glycosylation pattern in the phospholipase A 2 (PLA) of individual honeybees. Without reducing the disulfide bonds, PLA was digested with trypsin and filtered with a 3 kDa molecular weight (MW) cut‐off membrane. With this procedure, the glycopeptides could be isolated from the nonglycosylated peptides. After tryptic digestion and ultrafiltration, the disulfide bonds were reduced before analysis by CE. To reduce the adsorption, CE separation was performed on successive multiple ionic‐polymer (SMIL) polybrene (PB) coated capillary columns. The SMIL‐PB columns allowed partial separation of the glycopeptides and eight glycopeptides were identified by on‐line coupling of CE with electrospray ionization (ESI) mass spectrometry. The analysis of phospholipase A 2 from the venom of individual bees indicated that the variation and relative abundances of different glycopeptides were similar between the younger and the older bees. Copyright © 2000 John Wiley & Sons, Ltd.