Identification of the protein kinase A regulatory RIα-catalytic subunit interface by amide H/2H exchange and protein docking

摘要: An important goal after structural genomics is to build up the structures of higher-order protein–protein complexes from individual subunits. Often higher order are difficult obtain by crystallography. We have used an alternative approach in which catalytic (C) subunit and RIα regulatory (R) PKA were first subjected computational docking, top 100,000 solutions subsequently filtered based on amide hydrogen/deuterium (H/2H) exchange interface protection data. The resulting set forms ensemble which, besides inhibitor peptide binding site, a flat between C-terminal lobe C-subunit A- B-helices uniquely identified. This holoenzyme structure satisfies all previous experimental data complex allows prediction new contacts two