Apo-intermediate in the transport cycle of lactose permease (LacY)

作者：Madej M Gregor; Soro Sonya N; Kaback H Ronald^*

来源：Proceedings of the National Academy of Sciences of the United States of America, 2012, 109(44): E2970-E2978.

DOI：10.1073/pnas.1211183109

• membrane proteins
• modeling
• membrane transport
• conformational change

The lactose permease (LacY) catalyzes coupled stoichiometric symport of a galactoside and an H+. Crystal structures reveal 12, mostly irregular, transmembrane alpha-helices surrounding a cavity with sugar-and H+-binding sites at the apex, which is accessible from the cytoplasm and sealed on the periplasmic side (an inward-facing conformer). An outward-facing model has also been proposed based on biochemical and spectroscopic measurements, as well as the X-ray structure of a related symporter. Converging lines of evidence demonstrate that LacY functions by an alternating access mechanism. Here, we generate a model for an apo-intermediate of LacY based on crystallographic coordinates of LacY and the oligopeptide/H+ symporter. The model exhibits a conformation with an occluded cavity inaccessible from either side of the membrane. Furthermore, kinetic considerations and double electron-electron resonance measurements suggest that another occluded conformer with bound sugar exists during turnover. An energy profile for symport is also presented.

• 出版日期2012-10-30