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Title: Forces and dynamics of glucose and inhibitor binding to sodium glucose co-transporter SGLT1 studied by single molecule force spectroscopy
Authors: Isabel Neundlinger
Theeraporn Puntheeranurak
Linda Wildling
Christian Rankl
Lai Xi Wang
Hermann J. Gruber
Rolf K.H. Kinne
Peter Hinterdorfer
Johannes Kepler Universitat Linz
Mahidol University
Agilent Technologies Osterrreich GmbH
University of Maryland School of Medicine
Max Planck Institut fur molekulare Physiologie
Keywords: Biochemistry, Genetics and Molecular Biology
Issue Date: 1-Aug-2014
Citation: Journal of Biological Chemistry. Vol.289, No.31 (2014), 21673-21683
Abstract: Single molecule force spectroscopy was employed to investigate the dynamics of the sodium glucose co-transporter (SGLT1) upon substrate and inhibitor binding on the single molecule level. CHO cells stably expressing rbSGLT1 were probed by using atomic force microscopy tips carrying either thioglucose, 2′-aminoethyl β-D-glucopyranoside, or aminophlorizin. Poly(ethylene glycol) (PEG) chains of different length and varying end groups were used as tether. Experiments were performed at 10, 25 and 37 °C to address different conformational states of SGLT1. Unbinding forces between ligands and SGLT1 were recorded at different loading rates by changing the retraction velocity, yielding binding probability, width of energy barrier of the binding pocket, and the kinetic off rate constant of the binding reaction. With increasing temperature, width of energy barrier and average life time increased for the interaction of SGLT1 with thioglucose (coupled via acrylamide to a long PEG) but decreased for aminophlorizin binding. The former indicates that in the membrane-bound SGLT1 the pathway to sugar translocation involves several steps with different temperature sensitivity. The latter suggests that also the aglucon binding sites for transport inhibitors have specific, temperature-sensitive conformations. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.
ISSN: 1083351X
Appears in Collections:Scopus 2011-2015

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