The role of the unique motifs in the amino-terminal region of PKN on its enzymatic activity.
The yeast two-hybrid system and in vitro binding assay were carried out to characterize the interaction between the amino-terminal and carboxyl-terminal region of PKN. It was revealed that the amino-terminal region containing the regulatory domain associated with the carboxyl-terminal catalytic region. A synthetic peptide, corresponding to the amino acid residues of PKN from 39 to 53, with substitution of isoleucine46 with serine was shown to become a potent substrate for PKN, and its wild type synthetic peptide inhibited the phosphorylation by PKN. These results suggest that the amino-terminal region of PKN contains the pseudosubstrate sequence and acts as an autoinhibitory region.
Kitagawa
M
M
Department of Biology, Faculty of Science, Kobe University, Japan.
Shibata
H
H
Toshimori
M
M
Mukai
H
H
Ono
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D003001:Cloning, Molecular; D006801:Humans; D007700:Kinetics; D016350:Leucine Zippers; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry; D010766:Phosphorylation; D011493:Protein Kinase C; D017346:Protein Serine-Threonine Kinases; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae
D010455:Peptides; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C061570:protein kinase N; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; D011493:Protein Kinase C