Identification of an autoinhibitory domain in the insulin receptor tyrosine kinase |
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Authors: | Anna Filipek Thomas R Soderling |
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Institution: | (1) Vollum Institute for Advanced Biomedical Research, Oregon Health Sciences University, 3181 S.W. Sam Jackson Park Road, 97201 Portland, OR, USA;(2) Present address: Department of Muscle Biochemistry, Nencki Institute of Experimental Biology, 3 Pasteur St., 02-093 Warsaw, Poland |
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Abstract: | We have tested the hypothesis that activation of the insulin receptor tyrosine kinase is due to autophosphorylation of tyrosines 1146, 1150 and 1151 within a putative autoinhibitory domain. A synthetic peptide corresponding to residues 1134–1162, with tyrosines substituted by alanine or phenylalanine, of the insulin receptor subunit was tested for its inhibitory potency and specificity towards the tyrosine kinase activity. This synthetic peptide gave inhibition of the insulin receptor tyrosine kinase autophosphorylation and phosphorylation of the exogenous substrate poly(Glu, Tyr) with an approximate IC50 of 100 M. Inhibition appeared to be independent of the concentrations of insulin or the substrate poly(Glu, Tyr) but was decreased by increasing concentrations of ATP. This same peptide also inhibited the EGF receptor tyrosine kinase but not a serine/threonine protein kinase. These results are consistent with the hypothesis that this autophosphorylation domain contains an autoinhibitory sequence. (Mol Cell Biochem120: 103–110, 1993)Abbreviations IR
Insulin Receptor
- SDS/PAGE
Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis
- CaM
Calmodulin
- HEPES
4-(2-Hydroxyethyl)-Piperazineethane-Sulfonic Acid
- DMEM
Dulbecco's Modified Eagle' Medium
- PMSF
Phenylmethyl-Sulfonyl Fluoride
- HPLC
High Performance Liquid Chromatography
- PKC
Protein Kinase C
- PKI
Inhibitory Peptide for cAMP-Kinase
- CaMK II
Ca2+/Calmodulin-Dependent Protein Kinase II
- CaN A
A Subunit of Calcineurin |
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Keywords: | protein tyrosine kinase insulin receptor autoinhibitory domain |
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