title: Mapping of the SecA·SecY and SecA·SecG interfaces by site-directed in vivo photocross-linking.
abstract: The two major components of the Eubacteria Sec-dependent protein translocation system are the heterotrimeric channel-forming component SecYEG and its binding partner, the SecA ATPase nanomotor. Once bound to SecYEG, the preprotein substrate, and ATP, SecA undergoes ATP-hydrolytic cycles that drive the stepwise translocation of proteins. Although a previous site-directed in vivo photocross-linking study (Mori, H., and Ito, K. (2006) Proc. Natl. Acad. Sci. U.S.A. 103, 16159-16164) elucidated residues of SecY needed for interaction with SecA, no reciprocal study for SecA protein has been reported to date. In the present study we mapped residues of SecA that interact with SecY or SecG utilizing this approach. Our results show that distinct domains of SecA on two halves of the molecule interact with two corresponding SecY partners as well as with the central cytoplasmic domain of SecG. Our data support the in vivo relevance of the Thermotoga maritima SecA·SecYEG crystal structure that visualized SecYEG interaction for only one-half of SecA as well as previous studies indicating that SecA normally binds two molecules of SecYEG.
authors.affiliation: Department of Molecular Biology and Biochemistry, Wesleyan University, Middletown, Connecticut 06457, USA.
authors.forename: Sanchaita
authors.initials: S
authors.lastname: Das
authors.forename: Donald B
authors.initials: DB
authors.lastname: Oliver
mesh_terms: D001577:Benzophenones / Q000737:chemistry; D001692:Biological Transport; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017017:Methanococcus / Q000378:metabolism; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000737:chemistry; D010777:Photochemistry / Q000379:methods*; D011485:Protein Binding; D021381:Protein Transport; D020124:Thermotoga maritima / Q000378:metabolism
chemical_list: C488060:4-benzoylphenylalanine; D001577:Benzophenones; D026901:Membrane Transport Proteins; D010649:Phenylalanine