I) Facilitative Glucose Transport A) Facilitated diffusion different from passive diffusion 1) Diffusion faster than expected based on partition

coefficient 2) Saturated (a) As the glucose concentration in the EC medium increased, the rate of transport increased only up to a certain point (b) Further increases in glucose concentration dont affect transport rate 3) Specific to certain structurally related compounds (a) Glucose, galactose, mannose, but slower for other sugars like fructose (b) Stereospecific- D-Glucose transported faster than L-Glucose 4) Competitive inhibition similar compounds compete for biding site of transporter 5) Non competitive inhibition transport inhibited by structurally unrelated compounds (a) GLUT inhibited by cytochalasin B (binds to site on intracellular surface) 6) Regulated by hormones or other factors B) Structure of facilitative glucose transporters 1) First isolated from RBCs, found 4 related transporters, named GLUT1-GLUT5 (a) Present in liver, kidney, brain, intestine 2) Composed of 2-4 identical protein subunits (a) Heavily glycosylated integral membrane proteins (b) 12 TM helical regions (M1-M12) (c) Large intracellular loop btwn M^ and M7 (d) Large glycosylated EC loop btwn M! and M2 (e) Both N and C terminal ends are intracellular C) Mechanism of glucose transport 1) Transporter only has a single site for glucose binding (a) Can exist in 2 conformations (i) Glucose binding site exposed to either IC or EC medium 2) EC glucose binds, and protein alters conformation, now exposing glucose to IC medium 3) Glucose becomes unbound and delivered to cytoplasm (a) Protein can bind glucose at its IC surface and bring glucose back (b) Can spontaneously revert to original conformation with binding site on outside 4) Direction of transport determined b gradient of glucose concentration (a) Once glucose is inside, hexokinase phosphorylates it quickly (b) Phos glucose cant diffuse passively across PM and cant bind to transporter so glucose trapped inside cell (i) Glucose can be accumulated inside in form of glucose phosphate D) Insulin and diabetes 1) Produces a rapid increase in glucose transport, esp into muscle and fat cells (a) GLUT present in cells in vesicles in cytosol (b) Binding of insulin to receptor causes vesicles to fuse with PM and the transporters become integrated w/in PM 2) Diabetes is decrease in insulin production from decreased in number of B- cells which produce insulin in pancreas (a) Leads to decrease in glucose transport in fat/muscle tissue because a 50% decrease in transporter proteins w/in membrane II) Coupled Glucose Transport

A) Responsible for glucose absorption into body, found in intestine and proximal tubule of kidney, still similar to facilitative transporter 1) Increased rate, transport is saturated, specific, and inhibited (a) Inhibited by phlorizin 2) Difference is that transport is absolutely dependent on presence of Na+ (a) Na and glucose cotransported 3) Transport is electrogenic- transport produce the flow of current across PM (a) Should cause one mole of positive charge to be transported into the cell per mole of glucose of each sodium ion cotransported (b) Proved by recorded current across epithelium B) The energetics of Na+-glucose transport [ Na+ ] o [glu ] i 1) = ( 1) occurs in SGLT2 (one Na per one glucose) [ Na+ ] i [glu ] o [ Na + ] o 2 [glu ] i (2) occurs in SGLT1 (2 Na per glucose) 2) ( )= [ Na + ] i [glu ] o 3) larger differences in glucose concentration can be achieved if there is a potential difference across PM (a) b/c Na is positive, a negative potential drives Na into cell and increase ability of transporter to concentrate glucose w/in cytosol (b) greater potential difference, the larger the driving force for inward movement C) Structure of transporter and mechanism of transport 1) Similar to facilitative transporters, but no homology in AA sequence (a) Na-Glucose is 50-60 AA longer 2) 14 TM -helical regions with large extracellular loops, one is glycosylated (a) both N and C terminal are extracellular 3) Mechanisms of transport (a) 2 Na binding sites, and Na must bind fist on EC surface (b) binding of Na enhances binding of glucose (c) when both are bound, protein changes conformation that exposes biding site to interior and reduces affinity of glucose to protein (d) glucose and Na+ fall off and delivered to cell (e) protein spontaneously reverts to is previous configuration with both glucose and Na binding sites on the EC medium D) Glucose-galactose Malabsorption 1) rare recessively inherited defect in sugar metabolism (a) failure to absorb glucose or galactose, lactose (b) results in diarrhea until infant switches to milk lacking glucose, galactose, lactose 2) Caused by defect in Na-glucose contransporter (a) Missense mutation causes AA change which eliminates normal glucose and galactose transport