Issue 1, 2014

On the catalytic mechanism of polysaccharide lyases: evidence of His and Tyr involvement in heparin lysis by heparinase I and the role of Ca2+

Abstract

The structurally diverse polysaccharide lyase enzymes are distributed from plants to animals but share common catalytic mechanisms. One, heparinase I (F. heparinum), is employed in the production of the major anticoagulant drug, low molecular weight heparin, and is a mainstay of cell surface proteoglycan analysis. We demonstrate that heparinase I specificity and efficiency depend on the cationic form of the substrate. Ca2+–heparin, in which α-L-iduronate-2-O-sulfate residues adopt 1C4 conformation preferentially, is a substrate, while Na+–heparin is an inhibitor. His and Tyr residues are identified in the catalytic step and a model based on molecular dynamics and docking is proposed, in which deprotonated His203 initiates β-elimination by abstracting the C5 proton of the α-L-iduonate-2-O-sulfate residue in the substrate, and protonated Tyr357 provides the donor to the hexosamine leaving group.

Graphical abstract: On the catalytic mechanism of polysaccharide lyases: evidence of His and Tyr involvement in heparin lysis by heparinase I and the role of Ca2+

Supplementary files

Article information

Article type
Paper
Submitted
28 Aug 2013
Accepted
29 Oct 2013
First published
31 Oct 2013

Mol. BioSyst., 2014,10, 54-64

On the catalytic mechanism of polysaccharide lyases: evidence of His and Tyr involvement in heparin lysis by heparinase I and the role of Ca2+

C. R. Córdula, M. A. Lima, S. K. Shinjo, T. F. Gesteira, L. Pol-Fachin, V. J. Coulson-Thomas, H. Verli, E. A. Yates, T. R. Rudd, M. A. S. Pinhal, L. Toma, C. P. Dietrich, H. B. Nader and I. L. S. Tersariol, Mol. BioSyst., 2014, 10, 54 DOI: 10.1039/C3MB70370C

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Spotlight

Advertisements