Open AccessPhorbol ester binding to protein kinase C requires a cysteine-rich zinc-finger-like sequence.
Author(s) -
Yoshitaka Ono,
Tomoko Fujii,
Koichi Igarashi,
Toshiki Kuno,
Chikako Tanaka,
Ushio Kikkawa,
Yasutomi Nishizuka
Publication year - 1989
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.86.13.4868
Subject(s) - zinc finger , protein kinase c , biochemistry , binding domain , protein kinase a , biology , microbiology and biotechnology , cysteine , peptide sequence , conserved sequence , dna binding domain , binding protein , binding site , kinase , enzyme , gene , transcription factor
Protein kinase C normally has a tandem repeat of a characteristic cysteine-rich sequence in C1, the conserved region of the regulatory domain. These sequences resemble the DNA-binding zinc finger domain. For the gamma subspecies of rat brain protein kinase C, various deletion and point mutants in this domain were constructed, and the mutated proteins were expressed in Escherichia coli by using the T7 expression system. Radioactive phorbol 12,13-dibutyrate binding analysis indicated that a cysteine-rich zinc-finger-like sequence was essential for protein kinase C to bind phorbol ester and that one of two sequences was sufficient for the phorbol ester binding. Conserved region C2, another region in the regulatory domain, was apparently needed for the enzyme to require Ca2+ for phorbol ester binding activity.