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Title: {sup 1}H NMR spectroscopic studies establish that heparanase is a retaining glycosidase

Highlights: •{sup 1}H and {sup 13}C NMR chemical shifts of fondaparinux were fully assigned by 1D and 2D NMR techniques. •Hydrolysis of fondaparinux by heparanase was monitored by {sup 1}H NMR spectroscopy. •Heparanase is established to be a retaining glycosidase. -- Abstract: Heparanase is an endo-β-glucuronidase that cleaves heparan sulfate side chains of proteoglycans in basement membranes and the extracellular matrix (ECM). Heparanase is implicated in several diverse pathological processes associated with ECM degradation such as metastasis, inflammation and angiogenesis and is thus an important target for anti-cancer and anti-inflammatory drug discovery. Heparanase has been classed as belonging to the clan A glycoside hydrolase family 79 based on sequence analysis, secondary structure predictions and mutagenic analysis, and thus it has been inferred that it is a retaining glycosidase. However, there has been no direct experimental evidence to support this conclusion. Herein we describe {sup 1}H NMR spectroscopic studies of the hydrolysis of the pentasaccharide substrate fondaparinux by heparanase, and provide conclusive evidence that heparanase hydrolyses its substrate with retention of configuration and is thus established as a retaining glycosidase. Knowledge of the mechanism of hydrolysis may have implications for future design of inhibitors for this important drug target.
Authors:
 [1] ;  [2] ;  [1] ;  [2]
  1. Institute for Glycomics, Griffith University Gold Coast Campus, QLD 4222 (Australia)
  2. School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072 (Australia)
Publication Date:
OSTI Identifier:
22242244
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 443; Journal Issue: 1; Other Information: Copyright (c) 2013 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CARBON 13; CHEMICAL SHIFT; DRUGS; GLUCURONIDASE; GLYCOSIDES; HYDROLYSIS; INFLAMMATION; NEOPLASMS; NUCLEAR MAGNETIC RESONANCE; SPECTROSCOPY; STRUCTURAL CHEMICAL ANALYSIS; SUBSTRATES; SULFATES