Cellular munc18c levels can modulate glucose transport rate and GLUT4 translocation in 3T3L1 cells.
Munc18c has been shown to bind syntaxin 4 and to play a role in GLUT4 translocation and glucose transport, although this role is as yet poorly defined. In the present study, the effects of modulating the available level of munc18c on glucose transport and GLUT4 translocation were examined. Over-expression of munc18c in 3T3L1 adipocytes inhibited insulin-stimulated glucose transport by approximately 50%. Basal glucose transport rates were also decreased by approximately 25%. In contrast, microinjection of a munc18c polyclonal antibody stimulated GLUT4 translocation by approximately 60% over basal levels without affecting insulin-stimulated GLUT4 levels. Microinjection of a control antibody had no effect. These data are consistent with the likelihood that antibody microinjection sequesters munc18c enabling translocation/fusion of GLUT4 vesicles. Mutagenesis of a potential proline-directed kinase phosphorylation site in munc18c, T569, that in previous studies of its neuronal counterpart munc18a caused its dissociation from its complex with syntaxin 1a, had no effect on munc18c's association with syntaxin 4 or its inhibition of glucose transport, indicative that phosphorylation of this residue is not important for insulin regulation of glucose transport. The over-expression and microinjection sequestration data support an inhibitory role for munc18c on translocation/fusion of GLUT4 vesicles. They further show that altering the level of available munc18c in 3T3L1 cells can modulate glucose transport rates, indicating its potential as a target for therapeutics in diabetes.
Macaulay
S Lance
SL
CSIRO Health Sciences and Nutrition, 343 Royal Parade, Parkville, Vic. 3052, Australia. lance.macaulay@csiro.au
Grusovin
Julian
J
Stoichevska
Violet
V
Ryan
Joanne M
JM
Castelli
Laura A
LA
Ward
Colin W
CW
D016475:3T3 Cells; D017667:Adipocytes / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D000906:Antibodies / Q000008:administration & dosage; D001693:Biological Transport, Active; D003924:Diabetes Mellitus, Type 2 / Q000188:drug therapy / Q000378:metabolism; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism*; D051275:Glucose Transporter Type 4; D007328:Insulin / Q000494:pharmacology; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008845:Microinjections; D009004:Monosaccharide Transport Proteins / Q000378:metabolism*; D051938:Munc18 Proteins; D009124:Muscle Proteins; D016297:Mutagenesis, Site-Directed; D009419:Nerve Tissue Proteins; D010766:Phosphorylation; D011392:Proline / Q000737:chemistry; D011506:Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D050765:Qa-SNARE Proteins; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D050827:Syntaxin 1; D033921:Vesicular Transport Proteins
D000906:Antibodies; D051275:Glucose Transporter Type 4; D007328:Insulin; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D051938:Munc18 Proteins; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins; D011506:Proteins; D050765:Qa-SNARE Proteins; D011994:Recombinant Proteins; C495693:Slc2a4 protein, mouse; C494214:Stx1a protein, mouse; C497943:Stxbp3 protein, mouse; D050827:Syntaxin 1; D033921:Vesicular Transport Proteins; D011392:Proline; D005947:Glucose