RESUMEN
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.
Asunto(s)
Ratas , Ratones , Masculino , Humanos , Femenino , Animales , Adulto , Homología de Secuencia de Aminoácido , Análisis de Secuencia de ADN , Isoformas de Proteínas/genética , Fosforilación , Fosfoproteínas/genética , Datos de Secuencia Molecular , Perfilación de la Expresión Génica , ADN Complementario/química , Proteínas Quinasas Dependientes de AMP Cíclico/fisiología , Clonación Molecular , Línea Celular , Secuencia de Bases , Secuencia de AminoácidosRESUMEN
No abstract available.