Active site voltage clamp fluorometry of the sodium glucose cotransporter hSGLT1

Gorraitz, Edurne; Hirayama, Bruce A.; Paz, Adrian; Wright, Ernest M.; Loo, Donald D. F.

doi:10.1073/pnas.1713899114

Cited by 23 publications

(27 citation statements)

References 34 publications

(74 reference statements)

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“…We then rescored each of the top 10 binding poses, allowing the protein side chains within 4 Å of phlorizin to relax with the MM/GBSA 18 protocol. The top-ranked pose (green/red) occupies the same vestibule found for the MTS-TAMRA (2-((5(6)-tetramethyl-rhodamine)carboxylamino)ethyl methanethiosulfonate) reagent 19 and shows overlap of the phlorizin sugar moiety with the redocked glucose (transparent yellow/red), while the aglycon tail occupies the outer vestibule pointing towards the extracellular space (Fig. 1a, b ).…”

Section: Resultsmentioning

confidence: 69%

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Inhibitor binding mode and allosteric regulation of Na+-glucose symporters

Bisignano

Ghezzi

et al. 2018

Nat Commun

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Sodium-dependent glucose transporters (SGLTs) exploit sodium gradients to transport sugars across the plasma membrane. Due to their role in renal sugar reabsorption, SGLTs are targets for the treatment of type 2 diabetes. Current therapeutics are phlorizin derivatives that contain a sugar moiety bound to an aromatic aglycon tail. Here, we develop structural models of human SGLT1/2 in complex with inhibitors by combining computational and functional studies. Inhibitors bind with the sugar moiety in the sugar pocket and the aglycon tail in the extracellular vestibule. The binding poses corroborate mutagenesis studies and suggest a partial closure of the outer gate upon binding. The models also reveal a putative Na+ binding site in hSGLT1 whose disruption reduces the transport stoichiometry to the value observed in hSGLT2 and increases inhibition by aglycon tails. Our work demonstrates that subtype selectivity arises from Na+-regulated outer gate closure and a variable region in extracellular loop EL5.

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Section: Resultsmentioning

confidence: 69%

“…The results for wild-type hSGLT1 and hSGLT2 agree with previous reports 20 , 27 , 65 . Superscripts refer to data obtained for hSGLT1 mutants expressed in oocytes and reported previously 19 – 21 . Data reported without superscripts were generated for this study…”

Section: Resultsmentioning

confidence: 99%

Inhibitor binding mode and allosteric regulation of Na+-glucose symporters

Bisignano

Ghezzi

et al. 2018

Nat Commun

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“…These and other biophysical studies of human SGLT1 provide insights into the conformational changes associated with sodium binding and isomerisation of the transporter between the outward and inward facing structures (e.g. see [ 37 , 47 ]).…”

Section: Structure Of Sgltsmentioning

confidence: 99%

“…The location of transmembrane helices (TM) of the structural model of SGLT1 are shown (some helices have been removed for clarity), along with the boundary of the 600 Å 3 vestibule (blue area) bounded by the outer ends of TM1, TM2, TM3, TM6, TM9 and TM10. Reproduced from [ 47 ], distributed under the terms of the Creative Commons Attribution-NonCommerical-NoDerivatives International License 4.0 (CC BY-NC-ND; https://creativecommons.org/licenses/by-nc-nd/4.0/ ). This figure is available as part of a downloadable slideset …”

Section: Inhibitors Of Glucose Reabsorptionmentioning

confidence: 99%

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Physiology of renal glucose handling via SGLT1, SGLT2 and GLUT2

2018

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The concentration of glucose in plasma is held within narrow limits (4–10 mmol/l), primarily to ensure fuel supply to the brain. Kidneys play a role in glucose homeostasis in the body by ensuring that glucose is not lost in the urine. Three membrane proteins are responsible for glucose reabsorption from the glomerular filtrate in the proximal tubule: sodium−glucose cotransporters SGLT1 and SGLT2, in the apical membrane, and GLUT2, a uniporter in the basolateral membrane. ‘Knockout’ of these transporters in mice and men results in the excretion of filtered glucose in the urine. In humans, intravenous injection of the plant glucoside phlorizin also results in excretion of the full filtered glucose load. This outcome and the finding that, in an animal model, phlorizin reversed the symptoms of diabetes, has stimulated the development and successful introduction of SGLT2 inhibitors, gliflozins, in the treatment of type 2 diabetes mellitus. Here we summarise the current state of our knowledge about the physiology of renal glucose handling and provide background to the development of SGLT2 inhibitors for type 2 diabetes treatment.Electronic supplementary materialThe online version of this article (10.1007/s00125-018-4656-5) contains a slideset of the figures for download, which is available to authorised users.

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Sugar Transport Across Epithelia

Loo

Wright

2020

Physiology in Health and Disease

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Active site voltage clamp fluorometry of the sodium glucose cotransporter hSGLT1

Cited by 23 publications

References 34 publications

Inhibitor binding mode and allosteric regulation of Na+-glucose symporters

Inhibitor binding mode and allosteric regulation of Na+-glucose symporters

Physiology of renal glucose handling via SGLT1, SGLT2 and GLUT2

Sugar Transport Across Epithelia

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