Synthesis and phorbol ester-binding studies of the individual cysteine-rich motifs of protein kinase D.
To investigate the phorbol ester-binding properties of the individual cysteine-rich motifs of protein kinase D (PKD), the 52-mer peptides containing each cysteine-rich motif of PKD (PKD-C1A, PKD-C1B) have been synthesized. The [3H]phorbol-12,13-dibutyrate (PDBu) binding to PKD-C1A was affected drastically by incubation temperature while that to PKD-C1B was not. Scatchard analysis of [3H]PDBu binding to both PKD C1 peptides gave dissociation constants of 2.5 +/- 0.4 and 2.7 +/- 0.8 nM for PKD-C1A and PKD-C1B, respectively, indicating that the two cysteine-rich motifs of PKD are functionally equivalent like those of PKCgamma.
Irie
K
K
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Japan.
Nakahara
A
A
Ohigashi
H
H
Fukuda
H
H
Wender
P A
PA
Konishi
H
H
Kikkawa
U
U
D000595:Amino Acid Sequence; D003545:Cysteine / Q000737:chemistry*; D008969:Molecular Sequence Data; D015240:Phorbol 12,13-Dibutyrate / Q000378:metabolism*; D011485:Protein Binding; D011493:Protein Kinase C / Q000737:chemistry / Q000378:metabolism*
D015240:Phorbol 12,13-Dibutyrate; C089212:protein kinase D; D011493:Protein Kinase C; D003545:Cysteine