Differential phosphorylation of IRS-1 and IRS-2 by insulin and IGF-I receptors

Abstract

The specific contribution of insulin and IGF-I receptors to IRS-protein activation remains elusive. We studied the signalling properties of AspB10-insulin, an analog with enhanced affinity for the IGF-I receptor, in comparison to native insulin using primary human skeletal muscle cells. In myoblasts regular insulin and AspB10-insulin were equipotent in stimulating the IRS cascade, whereas this analog induced a significantly higher Shc phosphorylation. Phosphorylation of IRS-1 in response to insulin was inhibited equally by blocking either the insulin or the IGF-I receptor. IRS-1 activation by AspB10-insulin was only inhibited by blocking the IGF-I receptor. IRS-2 phosphorylation induced by both insulin and AspB10-insulin was nearly insensitive to blocking the insulin receptor, being predominantly mediated by the IGF-I receptor. We conclude that in myoblasts IRS-2, but not IRS-1, functions as preferred substrate for the IGF-I receptor. These data suggest a specific role for IRS-2 in growth and differentiation of human skeletal muscle.

Key words:  :

• IRS proteins
• insulin signalling
• myogenic differentiation
• myoblasts
• insulin receptor
• IGF-I receptor

Abbreviations
IRS	=	insulin receptor substrate
IGF-I	=	insulin-like growth factor
FCS	=	fetal calf serum
ECL	=	enhanced chemiluminescence
PBS	=	phosphate-buffered saline
TBS	=	Tris-buffered saline
SDS	=	sodium dodecyl sulphate
PAGE	=	polyacrylamide gel electrophoresis
BSA	=	bovine serum albumin
PBS	=	phosphate-buffered saline

Abbreviations
IRS	=	insulin receptor substrate
IGF-I	=	insulin-like growth factor
FCS	=	fetal calf serum
ECL	=	enhanced chemiluminescence
PBS	=	phosphate-buffered saline
TBS	=	Tris-buffered saline
SDS	=	sodium dodecyl sulphate
PAGE	=	polyacrylamide gel electrophoresis
BSA	=	bovine serum albumin
PBS	=	phosphate-buffered saline