Two negatively charged invariant residues influence ligand binding and conformational dynamics of 14-3-3ζ

FEBS Letters

article

Kruti Modi¹  Somavally Dalvi¹  Prasanna Venkatraman¹ 

[1] Protein Interactome Lab for Structural and Functional Biology, Advanced Centre for Treatment, Research and Education in Cancer;BARC Training School Complex, Homi Bhabha National Institute

关键词: 14-3-3 protein;    conformational change;    fluorescence anisotropy;    limited trypsinolysis;    peptide interaction;    surface plasmon resonance;   

DOI  :  10.1002/1873-3468.13662

来源: John Wiley & Sons Ltd.

【 摘 要 】

14-3-3 proteins bind and modulate the activities of a wide variety of phosphoproteins. Crystal structures of 14-3-3 isoforms bound to phospholigands have identified several residues important for ligand binding. Here, we report the role of two invariant residues, D124 and E131, in peptide binding and peptide-induced conformational changes of the binding pocket. Surprisingly, the D124A mutation abrogates peptide binding, while the E131A mutation results in a twofold increase in peptide affinity. The mutants are less stable than the wild-type protein, and peptide binding restores native-like stability to the E131A mutant. This reversibility is lost in the more open structure of D124A. Based on these results, we infer that E131 is a regulator of protein plasticity and D124 is the guardian of the active site geometry.

【 授权许可】

Unknown