| RRC ID |
72290
|
| Author |
Shindo M, Irie K, Ohigashi H, Kuriyama M, Saito N.
|
| Title |
Diacylglycerol kinase gamma is one of the specific receptors of tumor-promoting phorbol esters.
|
| Journal |
Biochem Biophys Res Commun
|
| Abstract |
Diacylglycerol kinase (DGK) and protein kinase C (PKC) are two different enzyme families that interact with diacylglycerol. Both enzymes contain cysteine-rich C1 domains with a zinc finger-like structure. Most of the C1 domains of PKCs show strong phorbol-12,13-dibutyrate (PDBu) binding with nanomolar dissociation constants (K(d)'s). However, there has been no experimental evidence that phorbol esters bind to the C1 domains of DGKs. We focused on DGK gamma because its C1A domain has a high degree of sequence homology to those of PKCs, and because DGK gamma translocates from the cytoplasm to the plasma membrane following 12-O-tetradecanoylphorbol-13-acetate treatment similar to PKCs. Two C1 domains of DGK gamma (DGK gamma-C1A and DGK gamma-C1B) were synthesized and tested for their PDBu binding along with whole DGK gamma (Flag-DGK gamma) expressed in COS-7 cells. DGK gamma-C1A and Flag-DGK gamma showed strong PDBu binding affinity, while DGK gamma-C1B was completely inactive. Scatchard analysis of DGK gamma-C1A and Flag-DGK gamma gave K(d)'s of 3.1 and 4.4 nM, respectively, indicating that the major PDBu binding site of DGK gamma is C1A. This is the first evidence that DGK gamma is a specific receptor of tumor-promoting phorbol esters.
|
| Volume |
289(2)
|
| Pages |
451-6
|
| Published |
2001-11-30
|
| DOI |
10.1006/bbrc.2001.5935
|
| PII |
S0006-291X(01)95935-2
|
| PMID |
11716494
|
| MeSH |
Amino Acid Sequence
Animals
CHO Cells
COS Cells
Cell Membrane / enzymology
Cricetinae
Cytoplasm / enzymology
Diacylglycerol Kinase / metabolism*
Dose-Response Relationship, Drug
Kinetics
Models, Genetic
Molecular Sequence Data
Mutation
Peptides / chemistry
Phorbol Esters / metabolism*
Plasmids / metabolism
Protein Binding
Protein Structure, Tertiary
Protein Transport
Sequence Homology, Amino Acid
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Spectrophotometry
Tetradecanoylphorbol Acetate / pharmacology
Zinc Fingers
|
| IF |
2.985
|
| Resource |
| Human and Animal Cells |
COS-7(RCB0539) |
