Expression, purification and characterization of the ubiquitous protein kinase C-related kinase 1
- 1 July 1995
- journal article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 309 (1) , 315-320
- https://doi.org/10.1042/bj3090315
Abstract
The recently described protein kinase C-related kinase (PRK) family is comprised of at least three members: PRK1, PRK2 and PRK3. Here the expression, purification and characterization of the ubiquitously expressed isoform, PRK1, is described. The enzyme was expressed in COS 7 cells and subsequently purified to apparent homogeneity by sequential column chromatography. The purified PRK1 protein migrates as a single 120 kDa polypeptide on SDS/PAGE. It displays a substrate specificity that in part resembles that of protein kinase C (PKC); however, unlike PKC, it is not activated by any combination of phorbol esters, diacylglycerol and Ca2+. Nevertheless, it can be activated by limited proteolysis, indicating a negative regulatory role for the N-terminal domain(s). PRK1 is also activated by phospholipids. The physiological relevance of this activation is discussed.Keywords
This publication has 14 references indexed in Scilit:
- Cloning and Expression Patterns of two Members of A Novel Protein‐kinase‐C‐related Kinase FamilyEuropean Journal of Biochemistry, 1995
- Antibodies to fluorylsulfonylbenzoyladenosine permit identification of protein kinasesFEBS Letters, 1993
- The regulatory domain of protein kinase C-ε restricts the catalytic-domain-specificityBiochemical Journal, 1991
- Purification and characterisation of bovine brain protein kinase C isotypes α, β and γEuropean Journal of Biochemistry, 1989
- Substrate-specific stimulation of protein kinase C by polyvalent anionsBiochemical and Biophysical Research Communications, 1987
- Activation of protein kinase C by Triton X-100 mixed micelles containing diacylglycerol and phosphatidylserine.Journal of Biological Chemistry, 1985
- Studies on a cyclic nucleotide-independent protein kinase and its proenzyme in mammalian tissues. II. Proenzyme and its activation by calcium-dependent protease from rat brain.Journal of Biological Chemistry, 1977
- Studies on a cyclic nucleotide-independent protein kinase and its proenzyme in mammalian tissues. I. Purification and characterization of an active enzyme from bovine cerebellum.Journal of Biological Chemistry, 1977
- 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