Lohmiller S, Hantke K, Patzer S I, Braun V
Max Planck Institute for Developmental Biology, Department of Protein Evolution, Tübingen, Germany.
Microbiology/Membrane Physiology, University of Tübingen, Germany.
Microbiology (Reading). 2008 Jun;154(Pt 6):1748-1754. doi: 10.1099/mic.0.2008/017350-0.
We have shown previously that Caulobacter crescentus grows on maltodextrins which are actively transported across the outer membrane by the MalA protein. Evidence for energy-coupled transport was obtained by deletion of the exbB exbD genes which abolished transport. However, removal of the TonB protein, which together with the ExbB ExbD proteins is predicted to form an energy-coupling device between the cytoplasmic membrane and the outer membrane, left transport unaffected. Here we identify an additional tonB gene encoded by the cc2334a ORF, which when deleted abolished maltose transport. MalA contains a TonB box that reads EEVVIT and is predicted to interact with TonB. Replacement of valine number 15 in the TonB box by proline abolished maltose transport. Maltose was transported across the cytoplasmic membrane by the MalY protein (CC2283). Maltose transport was induced by maltose and repressed by the MalI protein (CC2284). In addition to MalA, MalY and MalI, the mal locus encodes two predicted cytoplasmic alpha-amylases (CC2285 and CC2286) and a periplasmic glucoamylase (CC2282). The TonB dependence together with the previously described ExbB ExbD dependence demonstrates energy-coupled maltose transport across the outer membrane. MalY is involved in maltose transport across the cytoplasmic membrane by a presumably ion-coupled mechanism.
我们之前已经表明,新月柄杆菌能够利用麦芽糖糊精生长,麦芽糖糊精通过MalA蛋白被主动转运穿过外膜。通过缺失exbB exbD基因获得了能量偶联转运的证据,这消除了转运。然而,去除TonB蛋白(据预测,它与ExbB ExbD蛋白一起在细胞质膜和外膜之间形成能量偶联装置)并没有影响转运。在这里,我们鉴定出由cc2334a开放阅读框编码的另一个tonB基因,该基因缺失时会消除麦芽糖转运。MalA包含一个读码为EEVVIT的TonB框,预计它会与TonB相互作用。将TonB框中的第15位缬氨酸替换为脯氨酸会消除麦芽糖转运。麦芽糖通过MalY蛋白(CC2283)被转运穿过细胞质膜。麦芽糖转运由麦芽糖诱导,并受到MalI蛋白(CC2284)的抑制。除了MalA、MalY和MalI之外,mal基因座还编码两种预测的细胞质α淀粉酶(CC2285和CC2286)和一种周质葡糖淀粉酶(CC2282)。TonB依赖性以及先前描述的ExbB ExbD依赖性证明了能量偶联的麦芽糖跨外膜转运。MalY可能通过离子偶联机制参与麦芽糖跨细胞质膜的转运。