A Nonradioactive Assay for the Insulin Receptor Tyrosine Kinase: Use in Monitoring Receptor Kinase Activity after Activation of Overexpressed Protein Kinase Cα and High Glucose Treatment

1995 ◽  
Vol 231 (2) ◽  
pp. 323-332 ◽  
Author(s):  
Annegret Boge ◽  
Richard A. Roth
Keyword(s):  
Protein Kinase ◽  
Tyrosine Kinase ◽  
Insulin Receptor ◽  
Receptor Tyrosine Kinase ◽  
High Glucose ◽  
Kinase Activity ◽  
Receptor Kinase ◽  
Insulin Receptor Tyrosine Kinase ◽  
Protein Kinase Cα ◽  
Glucose Treatment

scholarly journals Effects of metformin on insulin receptor tyrosine kinase activity in rat adipocytes

Diabetologia ◽  
1986 ◽  
Vol 29 (11) ◽  
pp. 798-801 ◽  
Author(s):  
D. B. Jacobs ◽  
G. R. Hayes ◽  
J. A. Truglia ◽  
D. H. Lockwood
Keyword(s):  
Tyrosine Kinase ◽  
Insulin Receptor ◽  
Receptor Tyrosine Kinase ◽  
Kinase Activity ◽  
Tyrosine Kinase Activity ◽  
Insulin Receptor Tyrosine Kinase ◽  
Rat Adipocytes

scholarly journals Insulin receptor : tyrosine kinase activity and insulin action

1989 ◽  
Vol 29 (6) ◽  
pp. 653-661 ◽  
Author(s):  
R. Ballotti ◽  
Y. Le Marchand-Brustel ◽  
S. Gammeltoft ◽  
E. Van Obberghen
Keyword(s):  
Tyrosine Kinase ◽  
Insulin Receptor ◽  
Receptor Tyrosine Kinase ◽  
Insulin Action ◽  
Kinase Activity ◽  
Tyrosine Kinase Activity ◽  
Insulin Receptor Tyrosine Kinase

scholarly journals Antibodies to insulin receptor tyrosine kinase stimulate its activity towards exogenous substrates without inducing receptor autophosphorylation

1989 ◽  
Vol 260 (3) ◽  
pp. 749-756 ◽  
Author(s):  
V Baron ◽  
N Gautier ◽  
N Rochet ◽  
R Ballotti ◽  
B Rossi ◽  
...  
Keyword(s):  
Growth Factor ◽  
Tyrosine Kinase ◽  
Insulin Receptor ◽  
Receptor Tyrosine Kinase ◽  
Kinase Activity ◽  
Kinase Domain ◽  
Tyrosine Kinase Domain ◽  
Insulin Receptor Tyrosine Kinase ◽  
Substrate Phosphorylation ◽  
Exogenous Substrates

Anti-peptide antibodies directed against a highly-conserved sequence of the insulin receptor tyrosine kinase domain have been used to study the relationship between this specific region and kinase activation. Antibodies have been prepared by the injection into a rabbit of a synthetic peptide (P2) corresponding to residues 1110-1125 of the proreceptor. The peptide exhibits 88-95% sequence similarity with the corresponding sequence in the v-ros protein and in receptors for epidermal growth factor and for insulin-like growth factor 1. Two antibodies with different specificities could be separated from total antiserum obtained after immunization with P2. One antibody [anti-(P-Tyr)] cross-reacted with phosphotyrosine and immunoprecipitated solely autophosphorylated receptors. This antibody was shown to increase or decrease the receptor tyrosine kinase activity depending on its concentration. In all circumstances receptor autophosphorylation and substrate phosphorylation were modulated in a parallel fashion. The second antibody (anti-P2) failed to immunoprecipitate the insulin receptor, but was found to interact with both the peptide and the receptor by e.l.i.s.a. assay. Using a tyrosine co-polymer we found that anti-P2 activated the insulin receptor kinase leading to substrate phosphorylation at a level similar to that observed with insulin. This effect was additive to the hormonal effect. In contrast, receptor autophosphorylation was not modified by the anti-peptide. The differential effect of this anti-peptide further supports the idea that receptor autophosphorylation and kinase activity towards exogenous substrates might be independently regulated. Finally, our data suggest that conformational changes in the receptor tyrosine kinase domain may be sufficient for activation of its enzymic activity.

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