Identifying functionally important conformational changes in proteins: activation of the yeast α-factor receptor Ste2p.

ABSTRACT: We have developed a procedure in which disulfide cross-links are used to identify regions of proteins that undergo functionally important intramolecular motion. The approach was applied to the identification of disulfide bonds that stabilize the active state of the yeast α-mating pheromone receptor Ste2p, a member of the superfamily of G protein-coupled receptors. Cysteine residues were introduced at random positions in targeted regions of a starting allele of Ste2p that completely lacks cysteines. Libraries of mutated receptors were then screened for alleles that exhibit constitutive signaling. Two strongly activated alleles were recovered containing cysteine residues in transmembrane (TM) segments 5 and 6. Constitutive activity of these alleles was dependent on the presence of both intro

SUBMITTER: Taslimi A 

PROVIDER: S-EPMC3346263 | biostudies-literature | 2012 May

REPOSITORIES: biostudies-literature