title: On the role of the two extracytoplasmic substrate-binding domains in the ABC transporter OpuA.
abstract: Members of two transporter families of the ATP-binding cassette (ABC) superfamily use two or even four extracytoplasmic substrate-binding domains (SBDs) for transport. We report on the role of the two SBDs in the translocation cycle of the ABC transporter OpuA from Lactococcus lactis. Heterooligomeric OpuA complexes with only one SBD or one functional and one non-functional SBD (inactivated by covalent linkage of a substrate mimic) have been constructed, and the substrate binding and transport kinetics of the purified transporters, reconstituted in liposomes, have been determined. The data indicate that the two SBDs of OpuA interact in a cooperative manner in the translocation process by stimulating either the docking of the SBDs onto the translocator or the delivery of glycine betaine to the translocator. It appears that one of these initial steps, but not the later steps in translocation or resetting of the system to the initial state, is rate determining for transport. These new insights on the functional role of the extracytoplasmic SBDs are discussed in the light of the current knowledge of substrate-binding-protein-dependent ABC transporters.
authors: Department of Biochemistry, Groningen Biomolecular Science and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
authors: Esther
authors: E
authors: Biemans-Oldehinkel
authors: Bert
authors: B
authors: Poolman
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D001622:Betaine / Q000378:metabolism; D013294:Lactococcus lactis / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000502:physiology
chemical_list: D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D001622:Betaine; D000251:Adenosine Triphosphatases; C412743:OpuA protein, Lactococcus lactis