Synthesis of protein kinase Cdelta C1b domain by native chemical ligation methodology and characterization of its folding and ligand binding.
The C1b domain of protein kinase Cdelta (PKCdelta), a potent receptor for ligands such as diacylglycerol and phorbol esters, was synthesized by utilizing native chemical ligation. With this synthetic strategy, the domain was efficiently constructed and shown to have high affinity ligand binding and correct folding. The C1b domain has been utilized for the development of novel ligands for the control of phosphorylation by PKC family members. This strategy will pave the way for the efficient construction of C1b domains modified with fluorescent dyes, biotin, etc.
Ohashi
Nami
N
Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kandasurugadai, Chiyoda-ku, Tokyo 101-0062, Japan.
Nomura
Wataru
W
Kato
Mai
M
Narumi
Tetsuo
T
Lewin
Nancy E
NE
Blumberg
Peter M
PM
Tamamura
Hirokazu
H
D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D008024:Ligands; D010446:Peptide Fragments / Q000138:chemical synthesis; D011485:Protein Binding; D017510:Protein Folding; D051745:Protein Kinase C-delta / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary
D008024:Ligands; D010446:Peptide Fragments; D051745:Protein Kinase C-delta