Vol. 55 No. 2/2008, 269–280
on-line at: www.actabp.pl
Regular paper
Degenerate specificity of PDZ domains from RhoA-specific nucleotide exchange factors PDZRhoGEF and LARG
Katarzyna Smietana1, Monika Kasztura1, Marcin Paduch1, Urszula Derewenda2, Zygmunt S. Derewenda2 and Jacek Otlewski1
2Department 1Faculty
of Biotechnology, Laboratory of Protein Engineering, University of Wrocaw, Wrocaw, Poland; of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, USA
Received: 18 April, 2008; revised: 20 May, 2008; accepted: 29 May, 2008 available on-line: 07 June, 2008
PDZ domains are ubiquitous protein–protein interaction modules which bind short, usually carboxyterminal fragments of receptors, other integral or membrane-associated proteins, and occasionally cytosolic proteins. Their role in organizing multiprotein complexes at the cellular membrane is crucial for many signaling pathways, but the rules defining their binding specificity are still poorly understood and do not readily explain the observed diversity of their known binding partners. Two homologous RhoA-specific, multidomain nucleotide exchange factors PDZRhoGEF and LARG contain PDZ domains which show a particularly broad recognition profile, as suggested by the identification of five diverse biological targets. To investigate the molecular roots of this phenomenon, we constructed a phage display library of random carboxyterminal hexapeptides. Peptide variants corresponding to the sequences identified in library selection were synthesized and their affinities for both PDZ domains were measured and compared with those of peptides derived from sequences of natural partners. Based on the analysis of the binding sequences identified for PDZRhoGEF, we propose a sequence for an 'optimal' binding partner. Our results support the hypothesis that PDZ–peptide interactions may be best understood when one considers the sum of entropic and dynamic effects for each peptide as a whole entity, rather than preferences for specific residues at a given position.

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