SEPARATION AND ANALYSIS OF THE GLYCOFORM POPULATIONS OF RIBONUCLEASE-B USING CAPILLARY ELECTROPHORESIS

被引:84
作者
RUDD, PM
SCRAGG, IG
COGHILL, E
DWEK, RA
机构
[1] Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, South Parks Road
关键词
CAPILLARY ELECTROPHORESIS; GLYCOFORMS; OLIGOMANNOSE; RIBONUCLEASE-B;
D O I
10.1007/BF00731704
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The development of methods to separate, analyse and monitor changes in glycoform populations is essential if a more detailed understanding of the structure, function and processing of glycoproteins is to emerge. In this study, intact ribonuclease B was resolved by borate capillary electrophoresis into five populations according to the particular oligomannose structure associated with each glycoform. The relative proportions of these populations are correlated with the percentages obtained indirectly by analysis of the hydrazine released oligosaccharides using Bio-Gel P-4 gel filtration, matrix assisted laser desorption mass spectrometry and high performance anion exchange chromatography. Alterations in the composition of the glycoform populations during digestion of ribonuclease B with A. saitoi alpha(1-2)mannosidase were monitored by capillary electrophoresis (CE). Digestion of the free oligosaccharides under the same conditions, monitored by anion exchange chromatography, revealed a difference in rate, allowing some insight into the role of the protein during oligosaccharide processing. In conjunction with other methods, this novel application of CE may prove a useful addition to the techniques available for the study of glycoform populations.
引用
收藏
页码:86 / 91
页数:6
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