A synthetic peptide corresponding to the GLUT4 C-terminal cytoplasmic domain causes insulin-like glucose transport stimulation and GLUT4 recruitment in rat adipocytes.
In rat epididymal adipocytes, practically all of the major glucose transporter isoform GLUT4 is constitutively sequestered in intracellular membranes and moves to the plasma membrane in response to insulin, whereas about half of GLUT1, the minor isoform, is constitutively functional at the plasma membrane and thus less affected by insulin. Transfection studies using cells whose glucose transport is normally not regulated by insulin have suggested that the C-terminal cytoplasmic domain of GLUT4 is responsible for its constitutive intracellular sequestration. To test if this was also the case in a classical insulin target cell, we introduced synthetic peptides corresponding to the C-terminal cytoplasmic domain of GLUT4 and GLUT1 (GLUT4C and GLUT1C, respectively) into rat adipocytes and studied their effects on the glucose transport activity and the steady state GLUT4 and GLUT1 distribution between the plasma membrane and intracellular membranes in host cells. GLUT4C introduced into basal adipocytes caused a large (up to 4.5-fold) and dose-dependent increase in the plasma membrane GLUT4, with a proportional reduction in microsomal GLUT4, without affecting GLUT1 distribution. GLUT4C incorporation also caused a large (up to 3-fold) dose-dependent stimulation of 3-O-methyl D-glucose (3OMG) flux in host cells. GLUT4C caused little if any GLUT4 or GLUT1 redistribution and changes in 3OMG flux in insulin-stimulated adipocytes. GLUT1C, on the other hand, did not affect GLUT1 or GLUT4 targeting and 3OMG flux in host cells. These findings not only underscore the importance of the C-terminal cytoplasmic domain of GLUT4 in its constitutive intracellular sequestration in a classical insulin target cell but also suggest the existence of a regulatory protein in adipocytes that interacts with GLUT4 at its cytoplasmic domain, thus participating in the constitutive intracellular sequestration of GLUT4.
Lee
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Biophysics Laboratory, Veterans Administration Medical Center and the Department of Biophysical Sciences, State University of New York, Buffalo, New York 14215, USA.
Jung
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CY
D017667:Adipocytes / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000737:chemistry; D005947:Glucose / Q000378:metabolism*; D051272:Glucose Transporter Type 1; D051275:Glucose Transporter Type 4; D007328:Insulin / Q000494:pharmacology*; D007425:Intracellular Membranes / Q000378:metabolism; D008297:Male; D009004:Monosaccharide Transport Proteins / Q000737:chemistry / Q000378:metabolism*; D009124:Muscle Proteins; D010455:Peptides / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley
D051272:Glucose Transporter Type 1; D051275:Glucose Transporter Type 4; D007328:Insulin; D009004:Monosaccharide Transport Proteins; D009124:Muscle Proteins; D010455:Peptides; C495679:Slc2a1 protein, rat; C495694:Slc2a4 protein, rat; D005947:Glucose