Use of a nonpenetrating substrate analogue to study the molecular mechanism of the outer membrane dicarboxylate transport system in Escherichia coli K12.
We have recently demonstrated that a cell-surface dicarboxylate-binding protein (DBP) is involved in the outer membrane dicarboxylate transport system in Escherichia coli K12. The present report deals with our findings relating to the mode of action of this protein, and the identity and properties of the outer membrane integral protein which is involved in the translocation of dicarboxylic acids across the hydrophobic regions of the outer membrane. By the use of a nonpenetrating succinate analogue, aspartate-dextran, and through reconstitution studies with purified DBP, the cell-surface DBP is found to play an important role in succinate influx but not efflux. Transport studies with major outer membrane protein mutants indicate that the matrix protein (also referred to as protein I or porin) is the only outer membrane integral protein actively involved in the outer membrane dicarboxylate transport system. In the absence of a functional DBP, porin translocates succinate in a relatively less efficient and nonspecific manner. A tentative working model is proposed for this transport system. In this model, the cell-surface DBP is depicted as the substrate recognition component of the otherwise nonspecific porin channel. Together, this "DBP-porin channel complex" forms an efficient, specific transport channel for dicarboxylic acids.
D001692:Biological Transport / Q000187:drug effects; D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D003911:Dextrans / Q000494:pharmacology*; D029382:Dicarboxylic Acid Transporters; D003998:Dicarboxylic Acids / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism; D007700:Kinetics; D011392:Proline / Q000378:metabolism; D013386:Succinates / Q000378:metabolism*
D002352:Carrier Proteins; D003911:Dextrans; D029382:Dicarboxylic Acid Transporters; D003998:Dicarboxylic Acids; D013386:Succinates; C030334:aspartate dextran; D006639:Histidine; D011392:Proline