Understanding the basis of communication within protein domains is a major challenge in structural biology. We present structural and dynamical evidence for allosteric effects in a PDZ domain, PDZ2 from the tyrosine phosphatase PTP-BL, upon binding to a target peptide. The NMR structures of its free and peptide-bound states differ in the orientation of helix alpha 2 with respect to the remainder of the molecule, concomitant with a readjustment of the hydrophobic core. Using an ultrafast mixing instrument, we detected a deviation from simple bimolecular kinetics for the association with peptide that is consistent with a rate-limiting conformational change in the protein (k(obs) similar to 7 x 10(3) s(-1)) and an induced-fit model. Furthermore, the binding kinetics of 15 mutants revealed that binding is regulated by long-range interactions, which can be correlated with the structural rearrangements resulting from peptide binding. The homologous protein PSD-95 PDZ3 did not display a similar ligand-induced conformational change.

Demonstration of long-range interactions in a PDZ domain by NMR, kinetics, and protein engineering / Gianni, Stefano; Tine, Walma; Arcovito, Alessandro; Calosci, Nicoletta; Bellelli, Andrea; Åke, Engstrom; TRAVAGLINI ALLOCATELLI, Carlo; Brunori, Maurizio; Per, Jemth; Geerten W., Vuister. - In: STRUCTURE. - ISSN 0969-2126. - STAMPA. - 14:12(2006), pp. 1801-1809. [10.1016/j.str.2006.10.010]

Demonstration of long-range interactions in a PDZ domain by NMR, kinetics, and protein engineering

GIANNI, STEFANO;ARCOVITO, Alessandro;CALOSCI, NICOLETTA;BELLELLI, Andrea;TRAVAGLINI ALLOCATELLI, Carlo;BRUNORI, Maurizio;
2006

Abstract

Understanding the basis of communication within protein domains is a major challenge in structural biology. We present structural and dynamical evidence for allosteric effects in a PDZ domain, PDZ2 from the tyrosine phosphatase PTP-BL, upon binding to a target peptide. The NMR structures of its free and peptide-bound states differ in the orientation of helix alpha 2 with respect to the remainder of the molecule, concomitant with a readjustment of the hydrophobic core. Using an ultrafast mixing instrument, we detected a deviation from simple bimolecular kinetics for the association with peptide that is consistent with a rate-limiting conformational change in the protein (k(obs) similar to 7 x 10(3) s(-1)) and an induced-fit model. Furthermore, the binding kinetics of 15 mutants revealed that binding is regulated by long-range interactions, which can be correlated with the structural rearrangements resulting from peptide binding. The homologous protein PSD-95 PDZ3 did not display a similar ligand-induced conformational change.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11573/358307
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