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Characterization of the interaction between RhoA and the amino‐terminal region of PKN
Author(s) -
Shibata Hideki,
Mukai Hideyuki,
Inagaki Yoshimasa,
Homma Yoshimi,
Kimura Kazushi,
Kaibuchi Kozo,
Narumiya Shuh,
Ono Yoshitaka
Publication year - 1996
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(96)00385-7
Subject(s) - rhoa , amino acid , gtp' , binding site , small gtpase , in vitro , chemistry , plasma protein binding , biochemistry , gtpase , signal transduction , enzyme
The yeast two‐hybrid system and in vitro binding assay were carried out to characterize the interaction between PKN and a small GTP‐binding protein, RhoA. It was revealed that the region corresponding to the amino acid residues 33–111 in the amino‐terminal region of PKN was sufficient to confer the ability to associate with RhoA. Each synthetic peptide fragment corresponding to the amino acid residues 74–93 and 94–113 of PKN inhibited the interaction between PKN and RhoA in the in vitro binding assay, suggesting that this region is important in the association with RhoA. The endogenous and the GAP‐stimulated GTPase activity of RhoA was inhibited by the interaction with PKN, suggesting the presence of a regulatory mechanism that sustains the GTP‐bound active form of RhoA.