Demonstration of long-range interactions in a PDZ domain by NMR, kinetics, and protein engineering
Demonstration of long-range interactions in a PDZ domain by NMR, kinetics, and protein engineering
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DOI：
10.1016/j.str.2006.10.010
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发表时间：
2006-12-01
期刊：
影响因子：
5.7
通讯作者：
Vuister, Geerten W.
中科院分区：
文献类型：
--
作者：
Gianni, Stefano;Walma, Tine;Vuister, Geerten W.
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.
