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Biophys Rep. 2016;2(1):33-43. Epub 2016 Jul 1.

Combining biophysical methods to analyze the disulfide bond in SH2 domain of C-terminal Src kinase.

Author information

1
iHuman Institute, ShanghaiTech University, Shanghai, 201203 China ; Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461 USA.
2
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461 USA.

Abstract

The Src Homology 2 (SH2) domain is a structurally conserved protein domain that typically binds to a phosphorylated tyrosine in a peptide motif from the target protein. The SH2 domain of C-terminal Src kinase (Csk) contains a single disulfide bond, which is unusual for most SH2 domains. Although the global motion of SH2 domain regulates Csk function, little is known about the relationship between the disulfide bond and binding of the ligand. In this study, we combined X-ray crystallography, solution NMR, and other biophysical methods to reveal the interaction network in Csk. Denaturation studies have shown that disulfide bond contributes significantly to the stability of SH2 domain, and crystal structures of the oxidized and C122S mutant showed minor conformational changes. We further investigated the binding of SH2 domain to a phosphorylated peptide from Csk-binding protein upon reduction and oxidation using both NMR and fluorescence approaches. This work employed NMR, X-ray cryptography, and other biophysical methods to study a disulfide bond in Csk SH2 domain. In addition, this work provides in-depth understanding of the structural dynamics of Csk SH2 domain.

KEYWORDS:

C-terminal Src kinase; Disulfide bond; Nuclear magnetic resonance; Src homology 2

Conflict of interest statement

Dongsheng Liu and David Cowburn declare that they have no conflict of interest. Human and animal rights and informed consent This article does not contain any studies with human or animal subjects performed by any of the authors.

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