Specificity and action pattern of heparanase Bp, a β-glucuronidase from Burkholderia pseudomallei
No Thumbnail Available
Authors
Yu, Yanlei
Williams, Asher
Zhang, Xing
Fu, Li
Xia, Ke
Xu, Yongmei
Zhang, Fuming
Liu, Jian
Koffas, Mattheos
Linhardt, Robert J.
Issue Date
2019-08-01
Type
Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Alternative Title
Abstract
The specificity and action pattern of a β-glucuronidase derived from the pathogenic bacteria Burkholderia pseudomallei and expressed in Escherichia coli as a recombinant protein has been evaluated. While this enzyme shows activity on a number of glycosaminoglycans, our study has focused on its action on heparin, heparan sulfate and their biosynthetic intermediates as well as chemoenzymatically synthesized, structurally defined heparan sulfate oligosaccharides. These heparin/heparan sulfate (HP/HS) substrates examined varied in size and structure, but all contained an uronic acid (UA) residue β-(1→4) linked to a glucosamine residue. On the substrates tested, this enzyme (heparanase Bp) acted only on a glucuronic acid residue β-(1→4) linked to an N-acetylglucosamine, N-sulfoglucosamine or N-acetyl-6-O-sulfoglucosamine residue. A substrate was required to have a length of pentasaccharide or longer and heparanase Bp acted with a random endolytic action pattern on HP/HS. The specificity and glycohydrolase mechanism of action of heparanase Bp resembles mammalian heparanase and is complementary to the bacterial heparin lyases, which act through an eliminase mechanism on a glucosamine residue (1→4) linked to a UA residue, suggesting its utility as a tool for the structural determination of HP/HS as well as representing a possible model for the medically relevant mammalian heparanase. The utility heparanase Bp was demonstrated by the oligosaccharide mapping of heparin, which afforded resistant intact highly sulfated domains ranging from tetrasaccharide to >28-mer with a molecular weight >9000.
Description
Glycobiology, 29, 572–581
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Full Citation
Specificity and action pattern of heparanase Bp, a β-glucuronidase from Burkholderia pseudomallei, Y. Yu, A. Williams, X. Zhang, L. Fu, K. Xia, Y. Xu, F. Zhang, J. Liu, M. Koffas, R. J. Linhardt, Glycobiology, 29, 572–581, 2019.
Publisher
Terms of Use
Journal
Volume
Issue
PubMed ID
DOI
ISSN
14602423
9596658
9596658