Cellugyrin is a marker for a distinct population of intracellular Glut4-containing vesicles.
Although Glut4 traffic is routinely described as translocation from an "intracellular storage pool" to the plasma membrane, it has been long realized that Glut4 travels through at least two functionally distinct intracellular membrane compartments on the way to and from the cell surface. Biochemical separation and systematic studies of the individual Glut4-containing compartments have been limited by the lack of appropriate reagents. We have prepared a monoclonal antibody against a novel component protein of Glut4 vesicles and have identified this protein as cellugyrin, a ubiquitously expressed homologue of a major synaptic vesicle protein, synaptogyrin. By means of sucrose gradient centrifugation, immunoadsorption, and confocal microscopy, we have shown that virtually all cellugyrin is co-localized with Glut4 in the same vesicles. However, unlike Glut4, cellugyrin is not re-distributed to the plasma membrane in response to insulin stimulation, and at least 40-50% of the total population of Glut4 vesicles do not contain this protein. We suggest that cellugyrin represents a specific marker of a functionally distinct population of Glut4 vesicles that permanently maintains its intracellular localization and is not recruited to the plasma membrane by insulin.
Kupriyanova
T A
TA
Boston University School of Medicine, Boston, Massachusetts 02118, USA.
Kandror
K V
KV
D017667:Adipocytes / Q000166:cytology / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015415:Biomarkers / Q000032:analysis; D019556:COS Cells; D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D003594:Cytoplasmic Granules / Q000737:chemistry* / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects; D005455:Fluorescent Antibody Technique; D051275:Glucose Transporter Type 4; D007328:Insulin / Q000494:pharmacology; D008297:Male; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D008862:Microsomes, Liver / Q000378:metabolism; D009004:Monosaccharide Transport Proteins / Q000032:analysis* / Q000276:immunology; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D013347:Subcellular Fractions / Q000737:chemistry / Q000378:metabolism; D013395:Sucrose / Q000494:pharmacology; D062805:Synaptogyrins; D014162:Transfection
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D051275:Glucose Transporter Type 4; D007328:Insulin; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins; C495694:Slc2a4 protein, rat; D062805:Synaptogyrins; D013395:Sucrose