Ligand specificity modulated by prolyl imide bond Cis/Trans isomerization in the Itk SH2 domain: a quantitative NMR study

Patrick J Breheny; Alain Laederach; D Bruce Fulton; Amy H Andreotti

doi:10.1021/ja0375380

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Journal article

Ligand specificity modulated by prolyl imide bond Cis/Trans isomerization in the Itk SH2 domain: a quantitative NMR study

Patrick J Breheny, Alain Laederach, D Bruce Fulton and Amy H Andreotti

Journal of the American Chemical Society, Vol.125(51), pp.15706-15707

12/24/2003

DOI: 10.1021/ja0375380

PMID: 14677936

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Abstract

The Src homology 2 (SH2) domain of interleukin-2 tyrosine kinase (Itk) binds two separate ligands: a phosphotyrosine-containing peptide and the Itk Src homology 3 (SH3) domain. Binding specificity for these ligands is regulated via cis/trans isomerization of the Asn 286-Pro 287 imide bond in the Itk SH2 domain. In this study, we develop a novel method of analyzing chemical shift perturbation and cross-peak volumes to measure the affinities of both ligands for each SH2 conformer. We find that the cis imide bond containing SH2 conformer exhibits a 3.5-fold higher affinity for the Itk SH3 domain compared with binding of the trans conformer to the same ligand, while the trans conformer binds phosphopeptide with a 4-fold greater affinity than the cis-containing SH2 conformer. In addition to furthering the understanding of this system, the method presented here will be of general application in quantitatively determining the specificities of conformationally heterogeneous systems that use a molecular switch to regulate binding between multiple distinct ligands.

Nuclear Magnetic Resonance, Biomolecular - methods

Protein-Tyrosine Kinases - chemistry

Substrate Specificity

Ligands

Protein Conformation

Kinetics

src Homology Domains

Details

Title: Subtitle: Ligand specificity modulated by prolyl imide bond Cis/Trans isomerization in the Itk SH2 domain: a quantitative NMR study
Creators: Patrick J Breheny - Department of Biochemistry, Biophysics and Molecular Biology, Program of Bioinformatics and Computational Biology, Iowa State University, Ames, Iowa 50011, USA
Alain Laederach
D Bruce Fulton
Amy H Andreotti
Resource Type: Journal article
Publication Details: Journal of the American Chemical Society, Vol.125(51), pp.15706-15707
DOI: 10.1021/ja0375380
PMID: 14677936
NLM abbreviation: J Am Chem Soc
ISSN: 0002-7863
eISSN: 1520-5126
Publisher: United States
Grant note: AI43957 / NIAID NIH HHS
Language: English
Date published: 12/24/2003
Academic Unit: Biostatistics
Record Identifier: 9983997350502771

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