Disulfide Trapping for Modeling and Structure Determination of Receptor: Chemokine Complexes.

ABSTRACT: Despite the recent breakthrough advances in GPCR crystallography, structure determination of protein-protein complexes involving chemokine receptors and their endogenous chemokine ligands remains challenging. Here, we describe disulfide trapping, a methodology for generating irreversible covalent binary protein complexes from unbound protein partners by introducing two cysteine residues, one per interaction partner, at selected positions within their interaction interface. Disulfide trapping can serve at least two distinct purposes: (i) stabilization of the complex to assist structural studies and/or (ii) determination of pairwise residue proximities to guide molecular modeling. Methods for characterization of disulfide-trapped complexes are described and evaluated in terms of throughput,

SUBMITTER: Kufareva I 

PROVIDER: S-EPMC4782801 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature