80K-H Acts as a Signaling Bridge in Intact Living Cells Between PKCζ and the GLUT4 Translocation Regulator Munc18c

Abstract

Insulin triggers the translocation of glucose transporter GLUT4 to the plasma membrane. To understand the nature of the missing links between upstream insulin activated kinases and proteins of the GLUT4 translocation apparatus, the role of 80K-H was examined to test if it was one such missing link in live cells. Fluorescence correlation spectroscopy showed that the mobility of 80K-H was significantly decreased by insulin stimulation. This was dependent on the presence of PKCζ and an intact binding site for PKCζ. Insulin also increased the mobility of munc18c in an 80K-H- and PKCζ dependent manner. These results indicate that insulin induces dynamic associations between PKCζ, 80K-H, and munc18c and that 80K-H may act as a key signaling link between PKCζ and munc18c in live cells.

Key words::

• Insulin
• Glucose uptake
• Fluorescence correlation spectroscopy
• GLUT4
• Atypical PKC
• munc18c