Premium
NPC 15437 interacts with the C1 domain of protein kinase C An analysis using mutant PKC constructs
Author(s) -
Sullivan James P.,
Connor Jane R.,
Tiffany Carol,
Shearer Barry G.,
Burch Ronald M.
Publication year - 1991
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(91)80739-p
Subject(s) - protein kinase c , mutant , enzyme , phorbol , microbiology and biotechnology , kinase , cysteine , chemistry , protein kinase domain , enzyme activator , biochemistry , protein kinase a , biology , gene
We recently demonstrated that 2,6,diamino‐ N ‐([I‐(oxotridecyl)‐2‐piperidinyl]methyl)‐hexanamide (NPC 15437) is a selective inhibitor of PKC interacting at the regulatory domain of the enzyme. To further investigate the interaction of NPC 15437 with PKC we expressed a series of cDNAs encoding mutant PKC molecules in COS7 cells. NPC 15437 had no effect on the protein kinase activity of mutants lacking the N‐terminal region of the C1 domain. Further, NPC 15437 was a competitive inhibitor of the activation of PKCα by phorbol ester and attenuated the binding of phorbol ester to the enzyme in intact cells. The present study demonstrates that mutant enzyme constructs can be used to localize the site of interaction of NPC 15437 with PKC to residues 12–42, which encodes the pseudosubstrate binding domain and part of the first cysteine‐rich repeat sequence.