The goal of this project is to understand the molecular mechanism of intestinal glucose and sodium transport. The intestine is responsible for the absorption of dietary carbohydrate and more than 4 liters of fluid per day. The brush border Na/glucose cotransporter (SGLT1) is responsible for absorbing 180 grams of glucose and 115 grams of sodium and is also directly or indirectly involved in the absorption of water. Oral Rehydration Therapy, credited with saving thousands of infants a day from infectious diarrhea, is based on the coupling of glucose, salt and water transport by SGLT1. The goal of this project is to understand how human SGLT1 actively transports glucose from the gut into the intestinal epithelium across the brush border membrane. Our previous studies have determined that glucose transport is driven by the sodium and electrical gradients across the brush border and now we wish to explain how glucose and sodium transport are coupled. The human SGLT1 gene will be expressed in cultured cells and biophysical methods will be used to measure the kinetics of Na/glucose co transport. These methods include electrical and optical assays to determine both the steady-state kinetics and the conformational changes of the protein that underlie coupled transport. We have determined the crystal structure of SGLTs, identified a sugar and sodium binding sites, and obtained structural insight into the active transport. The structure will be used as a guide in our biophysical experiments to determine how glucose binds and is transported through the protein molecule. Our objective is to explain the kinetics and sugar selectivity of Na/glucose cotransport in terms of the structural dynamics of SGLT1. SGLT structures will provide clues about the differences in functional properties between the brush border Na/glucose cotransport and the glucosensor (SGLT3) found in the enteric nervous system. The results will also provide a structural basis for the interaction of natural products in our diet with SGLT1 and hopefully explain how these glycosides either blunt glucose absorption or how they are absorbed. Many such glycosides, components of folk medicines, are used to treat a variety of ailments including diabetes, obesity and aging. On a broader scale our work is also relevant to the function of other members of the SGLT gene family (SLC5), e.g. the Na/iodide and SGLT2, and the structurally related serotonin reuptake transporter (SERT) in brain and enteric nervous systems.

Public Health Relevance

This study is to determine how SGLTs transport glucose and sodium across the brush border membrane of the small intestine. The results of our studies may have a large impact on our understanding of glucose absorption in normal healthy subjects and the use of Oral Rehydration Therapy in the treatment of diarrhea. Our structural studies may also account for the differences in function between SGLT transporters and glucose sensors expressed in the GI tract and this will aid in the design of selective agonists and antagonists for the sensors. Some 21 SGLT drugs are currently in clinical trial for the treatment of diabetes. This will enhance therapies designed to regulate glucose absorption in diabetic and obese patients. Furthermore, our unexpected finding that the glucose and neurotransmitter transporters share a common structure has broad implications for transporters expressed in the enteric and central nervous systems.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK019567-34
Application #
8258781
Study Section
Gastrointestinal Cell and Molecular Biology Study Section (GCMB)
Program Officer
Grey, Michael J
Project Start
1976-12-01
Project End
2014-04-30
Budget Start
2012-05-01
Budget End
2013-04-30
Support Year
34
Fiscal Year
2012
Total Cost
$328,285
Indirect Cost
$115,113
Name
University of California Los Angeles
Department
Physiology
Type
Schools of Medicine
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
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Yu, Amy S; Hirayama, Bruce A; Timbol, Gerald et al. (2013) Regional distribution of SGLT activity in rat brain in vivo. Am J Physiol Cell Physiol 304:C240-7
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Sala-Rabanal, Monica; Hirayama, Bruce A; Loo, Donald D F et al. (2012) Bridging the gap between structure and kinetics of human SGLT1. Am J Physiol Cell Physiol 302:C1293-305
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Hummel, Charles S; Lu, Chuan; Loo, Donald D F et al. (2011) Glucose transport by human renal Na+/D-glucose cotransporters SGLT1 and SGLT2. Am J Physiol Cell Physiol 300:C14-21
Wright, Ernest M; Loo, Donald D F; Hirayama, Bruce A (2011) Biology of human sodium glucose transporters. Physiol Rev 91:733-94
Watanabe, Akira; Choe, Seungho; Chaptal, Vincent et al. (2010) The mechanism of sodium and substrate release from the binding pocket of vSGLT. Nature 468:988-91
Choe, Seungho; Rosenberg, John M; Abramson, Jeff et al. (2010) Water permeation through the sodium-dependent galactose cotransporter vSGLT. Biophys J 99:L56-8
Abramson, Jeff; Wright, Ernest M (2009) Structure and function of Na(+)-symporters with inverted repeats. Curr Opin Struct Biol 19:425-32

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