Hearn Elizabeth M, Patel Dimki R, Lepore Bryan W, Indic Mridhu, van den Berg Bert
Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
Nature. 2009 Mar 19;458(7236):367-70. doi: 10.1038/nature07678. Epub 2009 Feb 1.
Membrane proteins that transport hydrophobic compounds have important roles in multi-drug resistance and can cause a number of diseases, underscoring the importance of protein-mediated transport of hydrophobic compounds. Hydrophobic compounds readily partition into regular membrane lipid bilayers, and their transport through an aqueous protein channel is energetically unfavourable. Alternative transport models involving acquisition from the lipid bilayer by lateral diffusion have been proposed for hydrophobic substrates. So far, all transport proteins for which a lateral diffusion mechanism has been proposed function as efflux pumps. Here we present the first example of a lateral diffusion mechanism for the uptake of hydrophobic substrates by the Escherichia coli outer membrane long-chain fatty acid transporter FadL. A FadL mutant in which a lateral opening in the barrel wall is constricted, but which is otherwise structurally identical to wild-type FadL, does not transport substrates. A crystal structure of FadL from Pseudomonas aeruginosa shows that the opening in the wall of the beta-barrel is conserved and delineates a long, hydrophobic tunnel that could mediate substrate passage from the extracellular environment, through the polar lipopolysaccharide layer and, by means of the lateral opening in the barrel wall, into the lipid bilayer from where the substrate can diffuse into the periplasm. Because FadL homologues are found in pathogenic and biodegrading bacteria, our results have implications for combating bacterial infections and bioremediating xenobiotics in the environment.
转运疏水化合物的膜蛋白在多药耐药中发挥着重要作用,并可引发多种疾病,这凸显了蛋白质介导的疏水化合物转运的重要性。疏水化合物很容易分配到规则的膜脂双层中,而它们通过水性蛋白质通道的转运在能量上是不利的。针对疏水底物,已经提出了通过侧向扩散从脂双层获取的替代转运模型。到目前为止,所有被提出具有侧向扩散机制的转运蛋白都起外排泵的作用。在此,我们展示了大肠杆菌外膜长链脂肪酸转运蛋白FadL摄取疏水底物的侧向扩散机制的首个实例。一种FadL突变体,其桶壁上的侧向开口变窄,但在其他方面与野生型FadL结构相同,该突变体不能转运底物。铜绿假单胞菌FadL的晶体结构表明,β桶壁上的开口是保守的,并且勾勒出一条长的疏水通道,该通道可以介导底物从细胞外环境通过极性脂多糖层,并通过桶壁上的侧向开口进入脂双层,底物可以从脂双层扩散到周质中。由于在致病细菌和生物降解细菌中都发现了FadL同源物,我们的结果对于对抗细菌感染和环境中异生物的生物修复具有重要意义。