Molecular cloning and characterization of a new member of the RAC protein kinase family: association of the pleckstrin homology domain of three types of RAC protein kinase with protein kinase C subspecies and beta gamma subunits of G proteins

Biochem Biophys Res Commun. 1995 Nov 13;216(2):526-34. doi: 10.1006/bbrc.1995.2654.

Abstract

cDNA clones encoding the third member of the RAC protein kinase family, termed RAC-PK gamma, were isolated from a rat brain cDNA library. The deduced amino acid sequence of RAC-PK gamma was highly related to those of previously identified family members, RAC-PK alpha and beta, that have a pleckstrin homology domain and a protein-serine/threonine kinase catalytic domain at the amino- and carboxyl-terminal regions, respectively. Northern blot analysis indicated that RAC-PK gamma was expressed abundantly in brain and testis. Specific activities of RAC-PK alpha, beta, and gamma purified from transfected COS-7 cells were similar when measured by using myelin basic protein as a phosphate acceptor. Analysis using fusion proteins of glutathione S-transferase revealed that the pleckstrin homology domain of the three subtypes of RAC-PK associate with both protein kinase C subspecies and beta gamma subunits of G proteins. These results suggest that the pleckstrin homology domains of RAC protein kinase family could associate more than one protein to regulate the activity and/or intracellular distribution of this enzyme family by different ways.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blood Proteins / chemistry*
  • Brain / enzymology
  • Cell Line
  • Chlorocebus aethiops
  • Cloning, Molecular / methods
  • Consensus Sequence
  • Conserved Sequence
  • DNA, Complementary / metabolism
  • GTP-Binding Proteins / chemistry*
  • Gene Library
  • Macromolecular Substances
  • Molecular Sequence Data
  • Multigene Family
  • Organ Specificity
  • Phosphoproteins*
  • Polymerase Chain Reaction
  • Protein Kinase C / chemistry*
  • Protein-Serine-Threonine Kinases / biosynthesis*
  • Protein-Serine-Threonine Kinases / chemistry*
  • Protein-Serine-Threonine Kinases / isolation & purification
  • Proto-Oncogene Proteins c-akt
  • Rats
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Sequence Homology, Amino Acid

Substances

  • Blood Proteins
  • DNA, Complementary
  • Macromolecular Substances
  • Phosphoproteins
  • Recombinant Proteins
  • platelet protein P47
  • Protein-Serine-Threonine Kinases
  • Proto-Oncogene Proteins c-akt
  • Protein Kinase C
  • GTP-Binding Proteins

Associated data

  • GENBANK/D49836