CAP defines a second signalling pathway required for insulin-stimulated glucose transport.
Insulin stimulates the transport of glucose into fat and muscle cells. Although the precise molecular mechanisms involved in this process remain uncertain, insulin initiates its actions by binding to its tyrosine kinase receptor, leading to the phosphorylation of intracellular substrates. One such substrate is the Cbl proto-oncogene product. Cbl is recruited to the insulin receptor by interaction with the adapter protein CAP, through one of three adjacent SH3 domains in the carboxy terminus of CAP. Upon phosphorylation of Cbl, the CAP-Cbl complex dissociates from the insulin receptor and moves to a caveolin-enriched, triton-insoluble membrane fraction. Here, to identify a molecular mechanism underlying this subcellular redistribution, we screened a yeast two-hybrid library using the amino-terminal region of CAP and identified the caveolar protein flotillin. Flotillin forms a ternary complex with CAP and Cbl, directing the localization of the CAP-Cbl complex to a lipid raft subdomain of the plasma membrane. Expression of the N-terminal domain of CAP in 3T3-L1 adipocytes blocks the stimulation of glucose transport by insulin, without affecting signalling events that depend on phosphatidylinositol-3-OH kinase. Thus, localization of the Cbl-CAP complex to lipid rafts generates a pathway that is crucial in the regulation of glucose uptake.
Baumann
C A
CA
Department of Physiology, University of Michigan School of Medicine, Ann Arbor, Michigan 48109, USA.
Ribon
V
V
Kanzaki
M
M
Thurmond
D C
DC
Mora
S
S
Shigematsu
S
S
Bickel
P E
PE
Pessin
J E
JE
Saltiel
A R
AR
D016475:3T3 Cells; D000626:Aminopeptidases / Q000378:metabolism; D000818:Animals; D001692:Biological Transport; D051242:Caveolin 1; D022461:Caveolins; D002460:Cell Line; D010122:Cystinyl Aminopeptidase; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D051275:Glucose Transporter Type 4; D007328:Insulin / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D009004:Monosaccharide Transport Proteins / Q000378:metabolism; D009124:Muscle Proteins; D050722:Oncogene Protein v-cbl; D010766:Phosphorylation; D015682:Retroviridae Proteins, Oncogenic / Q000378:metabolism*; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques
C495541:Cav1 protein, mouse; D051242:Caveolin 1; D022461:Caveolins; D003598:Cytoskeletal Proteins; D051275:Glucose Transporter Type 4; D007328:Insulin; D008563:Membrane Lipids; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D009124:Muscle Proteins; D050722:Oncogene Protein v-cbl; D015682:Retroviridae Proteins, Oncogenic; C495693:Slc2a4 protein, mouse; C106354:flotillins; D000626:Aminopeptidases; D010122:Cystinyl Aminopeptidase; C105964:leucyl-cystinyl aminopeptidase; D005947:Glucose