Mapping of the ligand-binding site on the b' domain of human PDI: interaction with peptide ligands and the x-linker region

Byrne, Lee J. and Sidhu, Ateesh and Wallis, Anne Katrine and Ruddock, Lloyd W. and Freedman, Robert B. and Howard, Mark J. and Williamson, Richard A. (2009) Mapping of the ligand-binding site on the b' domain of human PDI: interaction with peptide ligands and the x-linker region. Biochemical Journal, 423 (2). pp. 209-217. ISSN 0264-6021. (Full text available)

PDF
Download (809kB)
[img]
Preview
Official URL
http://dx.doi.org/10.1042/BJ20090565

Abstract

PDI (protein disulfide-isomerase) catalyses the formation of native disulfide bonds of secretory proteins in the endoplasmic reticulum. PDI consists of four thioredoxin-like domains, of which two contain redox-active catalytic sites (a and a'), and two do not (b and b'). The b' domain is primarily responsible for substrate binding, although the nature and specificity of the substrate-binding site is still poorly understood. In the present study, we show that the b' domain of human PDI is in conformational exchange, but that its structure is stabilized by the addition of peptide ligands or by binding the x-linker region. The location of the ligand-binding site in b' was mapped by NMR chemical shift perturbation and found to consist primarily of residues from the core beta-sheet and alpha-helices 1 and 3. This site is where the x-linker region binds in the X-ray structure of b'x and we show that peptide ligands can compete with x binding at this site. The finding that x binds in the principal ligand-binding site of b' further supports the hypothesis that x functions to gate access to this site and so modulates PDI activity.

Item Type: Article
Additional information: Byrne, Lee J Sidhu, Ateesh Wallis, A Katrine Ruddock, Lloyd W Freedman, Robert B Howard, Mark J Williamson, Richard A BB/D018072/Biotechnology and Biological Sciences Research Council/United Kingdom Research Support, Non-U.S. Gov't England The Biochemical journal Biochem J. 2009 Sep 25;423(2):209-17.
Uncontrolled keywords: Binding Sites Humans Ligands Models, Molecular Peptide Fragments/chemistry/isolation & purification/*metabolism Protein Disulfide-Isomerases/*chemistry/isolation & purification/*metabolism *Protein Interaction Mapping Protein Structure, Secondary Protein Structure, Tertiary/physiology Somatostatin/chemistry/metabolism
Subjects: Q Science > QD Chemistry
Q Science > QH Natural history > QH301 Biology
Divisions: Faculties > Science Technology and Medical Studies > School of Biosciences > Protein Science Group
Depositing User: Mark Howard
Date Deposited: 16 Aug 2011 03:56
Last Modified: 18 Jul 2014 13:30
Resource URI: http://kar.kent.ac.uk/id/eprint/28011 (The current URI for this page, for reference purposes)
  • Depositors only (login required):

Downloads

Downloads per month over past year