Activation of PKN, a novel 120-kDa protein kinase with leucine zipper-like sequences, by unsaturated fatty acids and by limited proteolysis

H Mukai; M Kitagawa; H Shibata; H Takanaga; K Mori; M Shimakawa; M Miyahara; K Hirao; Y Ono

doi:10.1006/bbrc.1994.2466

Activation of PKN, a novel 120-kDa protein kinase with leucine zipper-like sequences, by unsaturated fatty acids and by limited proteolysis

Biochem Biophys Res Commun. 1994 Oct 14;204(1):348-56. doi: 10.1006/bbrc.1994.2466.

Authors

H Mukai¹, M Kitagawa, H Shibata, H Takanaga, K Mori, M Shimakawa, M Miyahara, K Hirao, Y Ono

Affiliation

¹ Department of Biology, Faculty of Science, Kobe University, Japan.

PMID: 7945381
DOI: 10.1006/bbrc.1994.2466

Abstract

PKN, a novel protein kinase with catalytic domain homologous to PKC family and unique amino terminal leucine zipper-like sequences, was purified partially from COS7 cells transfected with the cDNA construct encoding human PKN for enzymatic characterization of the enzyme. Using serine containing synthetic peptides based on PKC pseudosubstrate sites as the phosphate acceptors, kinase activities estimated from partially purified PKN were not stimulated by Ca2+/phosphatidylserine/diolein but were activated several-fold to several tens-fold by 40 microM unsaturated fatty acids, such as arachidonic acid, linoleic acid, and oleic acid. Autophosphorylation of the immunoprecipitates using anti-PKN antiserum was also stimulated by various unsaturated fatty acids. Limited proteolysis of PKN with trypsin induced an enhancement of the peptide kinase activity that was almost independent of arachidonic acid.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Cell Line
Chlorocebus aethiops
Chromatography, Ion Exchange
Enzyme Activation
Fatty Acids, Nonesterified / pharmacology
Fatty Acids, Unsaturated / pharmacology*
Humans
Kidney
Kinetics
Leucine Zippers
Molecular Sequence Data
Peptide Fragments / chemistry
Peptide Fragments / isolation & purification
Phosphorylation
Protein Kinase C
Protein Kinases / biosynthesis
Protein Kinases / isolation & purification
Protein Kinases / metabolism*
Protein Serine-Threonine Kinases*
Protein-Tyrosine Kinases / biosynthesis
Protein-Tyrosine Kinases / isolation & purification
Protein-Tyrosine Kinases / metabolism*
Recombinant Proteins / biosynthesis
Recombinant Proteins / isolation & purification
Recombinant Proteins / metabolism
Substrate Specificity
Transfection
Trypsin

Substances

Fatty Acids, Nonesterified
Fatty Acids, Unsaturated
Peptide Fragments
Recombinant Proteins
Protein Kinases
protein kinase N
Protein-Tyrosine Kinases
Protein Serine-Threonine Kinases
Protein Kinase C
Trypsin