Liquid Chromatography-Mass Spectrometry to Study Chondroitin Lyase Action Pattern

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Authors
Zhang, Zhenqing
Park, Youmie
Kemp, Melissa M.
Zhao, Wenjing
Im, A. Rang
Shaya, David
Cygler, Miroslaw
Kim, Yeong Shik
Linhardt, Robert J.
Issue Date
2009-02-01
Type
Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
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Abstract
Liquid chromatography–mass spectrometry was applied to determine the action pattern of different chondroitin lyases. Two commercial enzymes, chondroitinase ABC (Proteus vulgaris) and chondroitinase ACII (Arthrobacter aurescens), having action patterns previously determined by viscosimetry and gel electrophoresis were first examined. Next, the action patterns of recombinant lyases, chondroitinase ABC from Bacteroides thetaiotaomicron (expressed in Escherichia coli) and chondroitinase AC from Flavobacterium heparinum (expressed in its original host), were examined. Chondroitin sulfate A (CS-A, also known as chondroitin-4-sulfate) was used as the substrate for these four lyases. Aliquots taken at various time points were analyzed. The products of chondroitinase ABC (P. vulgaris) and chondroitinase AC (F. heparinum) contained unsaturated oligosaccharides of sizes ranging from disaccharide to decasaccharide, demonstrating that both are endolytic enzymes. The products afforded by chondroitinase ABC (B. thetaiotaomicron) and chondroitinase ACII (A. aurescens) contained primarily unsaturated disaccharide. These two exolytic enzymes showed different minor products, suggesting some subtle specificity differences between the actions of these two exolytic lyases on chondroitin sulfate A.
Description
Analytical Biochemistry, 385, 57-64
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Full Citation
Liquid Chromatography-Mass Spectrometry to Study Chondroitin Lyase Action Pattern, Z. Zhang, Y. Park, M. M. Kemp, W. Zhao, A.-R. Im, D. Shaya, M. Cygler, Y. S. Kim, R. J. Linhardt, Analytical Biochemistry, 385, 57-64, 2008.
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10960309
32697
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