Phosphoryl exchange is involved in the mechanism of the insulin receptor kinase
- 25 July 1994
- journal article
- Published by Wiley in FEBS Letters
- Vol. 349 (1) , 17-22
- https://doi.org/10.1016/0014-5793(94)00632-6
Abstract
The cytoplasmic kinase domain of the human insulin receptor (IRKD; M r 49 kDa) has been over‐expressed in insect cells using the baculovirus expression system. To investigate the kinase mechanism, we have compared the stoichiometry of ADP formation and phosphoryl transfer. After an initial phase of autophosphorylation, ATP is consumed without a stoichiometric increase in incorporated phosphate. During substrate phosphorylation using poly(Glu: Tyr) (4:1) phosphoryl transfer comes close to ATP turnover, which is independent of the presence of the substrate, indicating an increased efficiency (i.e. ATP turnover/phosphate incorporation) of phosphoryl transfer. Autophosphorylation under pulse‐chase conditions suggests the existence of a phosphoenzyme intermediate.Keywords
This publication has 20 references indexed in Scilit:
- Insulin receptor autophosphorylation. I. Autophosphorylation kinetics of the native receptor and its cytoplasmic kinase domainBiochemistry, 1993
- Signal transduction by receptors with tyrosine kinase activityPublished by Elsevier ,1990
- [19] Enzymatic and chemical digestion of proteins for mass spectrometryPublished by Elsevier ,1990
- Another version of the human insulin receptor kinase domain: expression, purification, and characterization.Proceedings of the National Academy of Sciences, 1989
- Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDaAnalytical Biochemistry, 1987
- Protein kinase assay by paper-trichloroacetic acid method: High performance using phosphocellulose paper and washing an ensemble of samples on flat sheetsAnalytical Biochemistry, 1987
- Human insulin receptor and its relationship to the tyrosine kinase family of oncogenesNature, 1985
- A modified high-performance thin-layer plate for the separation of purines and pyrimidinesAnalytical Biochemistry, 1983
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970