Exp Mol Med.
2006 Apr;38(2):144-152.
Protein kinase A-dependent phosphorylation of B/K protein
- Affiliations
-
- 1National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.
- 2Department of Biochemistry, College of Medicine, The Catholic University of Korea, Seoul 137-701, Korea. ojkwon@catholic.ac.kr
- 3R and D center, PetaGen Inc., B199F Yonsei Engineering Research Complex, Seoul 120-140, Korea.
- 4Princeton University, Princeton NJ 08544, USA.
Abstract
- We have previously isolated a novel protein "B/K" that contains two C2-like domains. Here, we report the isolatioin and mRNA distribution of a human B/K isoform, and protein kinase A (PKA)-dependent phosphorylation of the B/K protein. The 1.5 kb human B/K cDNA clone exhibits 89% and 97% identities with rat B/K in the sequences of nucleotide and amino acid, respectively. Human B/K isoform encodes a 474 amino acid protein and shows structural features similar to the rat counterpart including two C2 domains, three consensus sequences for PKA, absence of a transmembrane region, and conservation of the N-terminal cysteine cluster. On Northern and dot blot analyses, a 3.0 kb B/K transcript was abundantly present in human brain, kidney, and prostate. Among the brain regions, strong signals were observed in the frontal and temporal lobes, the hippocampus, the hypothalamus, the amygdala, the substantia nigra, and the pituitary. Recombinant B/K proteins containing three consensus sites for PKA was very efficiently phosphorylated in vitro by PKA catalytic subunit. B/K protein which was overexpressed in LLC-PK1 cells was also strongly phosphorylated in vivo by vasopressin analog DDAVP, and PKA-specific inhibitor H89 as well as type 2 vasopressin receptor antagonist specifically suppressed DDAVP-induced B/K phosphorylation. These results suggest that B/K proteins play a role as potential substrates for PKA in the area where they are expressed.
MeSH Terms
-
Sequence Homology, Amino Acid
Sequence Analysis, DNA
Rats
Protein Isoforms/genetics
Phosphorylation
Phosphoproteins/genetics/*metabolism
Molecular Sequence Data
Mice
Male
Humans
Gene Expression Profiling
Female
DNA, Complementary/chemistry/genetics
Cyclic AMP-Dependent Protein Kinases/*physiology
Cloning, Molecular
Cell Line
Base Sequence
Animals
Amino Acid Sequence
Adult
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