Synthesis and phorbol ester-binding studies of the individual cysteine-rich motifs of protein Kinase D

• Published in: Bioorganic & medicinal chemistry letters 1999-09, Vol.9 (17), p.2487-2490
• Main Authors: Irie, Kazuhiro, Nakahara, Akifumi, Ohigashi, Hajime, Fukuda, Hiroyuki, Wender, Paul A., Konishi, Hiroaki, Kikkawa, Ushio
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Cysteine - chemistry; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Molecular Sequence Data; Phorbol 12,13-Dibutyrate - metabolism; Protein Binding; Protein Kinase C - chemistry; Protein Kinase C - metabolism; Transferases
• ISSN: 0960-894X; 1464-3405
• DOI: 10.1016/S0960-894X(99)00413-8

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 PKCγ.