Engineered occluded apo-intermediate of LacY

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

The lactose permease (LacY) catalyzes coupled stoichiometric symport of a galactoside and an H(+). Crystal structures reveal 12, mostly irregular, transmembrane α-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...

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Evidence for an intermediate conformational state of LacY.

LacY mutant Cys154 → Gly exhibits a periplasmic-closed crystal structure identical to the WT, but is periplasmic-open in the membrane. The mutant hardly catalyzes transport, but binds galactosides from either side of the membrane with the same affinity and is resistant to site-directed proteolysis relative to the pseudo-WT. Site-directed alkylation was also applied to 11 single-Cys mutants in C...

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Flexible Gates Generate Occluded Intermediates in the Transport Cycle of LacY☆

The major facilitator superfamily (MFS) transporter lactose permease (LacY) alternates between cytoplasmic and periplasmic open conformations to co-transport a sugar molecule together with a proton across the plasma membrane. Indirect experimental evidence suggested the existence of an occluded transition intermediate of LacY, which would prevent leaking of the proton gradient. As no experiment...

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Intermediate view synthesis considering occluded and ambiguously referenced image regions

In this paper, we present an algorithm for synthesizing intermediate views from a single stereopair. The key contributions of this algorithm are the incorporation of scene assumptions and a disparity estimation confidence measure that lead to the accurate synthesis of occluded and ambiguously referenced regions. The synthesized views have been displayed on a multi-view binocular imaging system,...

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Electrophysiological characterization of LacY.

Electrogenic events due to the activity of wild-type lactose permease from Escherichia coli (LacY) were investigated with proteoliposomes containing purified LacY adsorbed on a solid-supported membrane electrode. Downhill sugar/H(+) symport into the proteoliposomes generates transient currents. Studies at different lipid-to-protein ratios and at different pH values, as well as inactivation by N...

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ژورنال

عنوان ژورنال: Proceedings of the National Academy of Sciences

سال: 2018

ISSN: 0027-8424,1091-6490

DOI: 10.1073/pnas.1816267115