The glucose transporter GluT4 and secretory carrier membrane proteins (SCAMPs) colocalize in rat adipocytes and partially segregate during insulin stimulation.
Secretory carrier membrane proteins (SCAMPs) mark the recycling system for the insulin-responsive glucose transporter, GluT4, in rat adipocytes. Anti-GluT4 and anti-SCAMP antibodies each immunoadsorbed vesicles containing both antigens from a low density microsomal fraction that is enriched in both antigens. The immunoadsorbed vesicles also contain VAMPs (synaptobrevins), synaptic vesicle membrane proteins. All three antigens were colocalized in low density microsomal vesicles from both basal and insulin-stimulated adipocytes. The SCAMPs have the same electrophoretic mobility as a major polypeptides detected in GluT4 vesicles. During insulin stimulation, 40% each of GluT4 and VAMPs redistribute from low density microsomes to the plasma membrane fraction; however, < 10% of the SCAMPs redistribute. Immunocytochemical staining of adipose tissue shows almost complete coincidence of SCAMPs and GluT4 in the basal state and extensive redistribution of both antigens to the cell periphery during insulin stimulation. Segregation of antigens during stimulation is not as distinct as observed by fractionation, although there are regions at the cell border where the SCAMPs appear more concentrated than GluT4. These data suggest that during insulin stimulation, in contrast to the behaviour of GluT4, SCAMPs remain tightly associated with the recycling system.
Laurie
S M
SM
Department of Anatomy and Cell Biology, University of Virginia Health Sciences Center, Charlottesville 22908.
Cain
C C
CC
Lienhard
G E
GE
Castle
J D
JD
D000273:Adipose Tissue / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D002352:Carrier Proteins / Q000032:analysis / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D005455:Fluorescent Antibody Technique; D051275:Glucose Transporter Type 4; D015151:Immunoblotting; D007150:Immunohistochemistry; D007163:Immunosorbent Techniques; D007328:Insulin / Q000494:pharmacology*; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D008854:Microscopy, Electron; D008861:Microsomes / Q000737:chemistry; D009004:Monosaccharide Transport Proteins / Q000032:analysis / Q000378:metabolism*; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins / Q000378:metabolism; D050683:R-SNARE Proteins; D051381:Rats; D017207:Rats, Sprague-Dawley
D002352:Carrier Proteins; D051275:Glucose Transporter Type 4; D007328:Insulin; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins; D050683:R-SNARE Proteins; C495694:Slc2a4 protein, rat