Involvement of beta-adrenergic receptor kinase-1 in homologous desensitization of histamine H2 receptors in human gastric carcinoma cell line MKN-45.
The poorly differentiated human gastric carcinoma cell line MKN-45 possesses histamine H2 receptors which are homologously desensitized by histamine. In order to clone the cDNA of a receptor kinase specific for H2 receptors, we performed RT-PCR at a low annealing temperature using oligo-DNA primers bearing the conserved sequences of the kinase domain of the G protein-coupled receptor kinase (GRK) family. However, we were able to isolate only cDNAs for beta-adrenergic receptor kinase 1 (beta ARK1) and GRK6. Interestingly, treatment of MKN-45 cells with beta ARK1 antisense phosphorothioate oligo-DNA (PON) resulted in significant loss of desensitization of H2 receptors by histamine, whereas GRK6 antisense PON had no effect. Thus, beta ARK1 appears to be involved in the homologous desensitization of H2 receptors in MKN-45 cells.
Nakata
H
H
Department of Internal Medicine, Kobe University School of Medicine, Japan.
Kinoshita
Y
Y
Kishi
K
K
Fukuda
H
H
Kawanami
C
C
Matsushima
Y
Y
Asahara
M
M
Okada
A
A
Maekawa
T
T
Chiba
T
T
D015152:Blotting, Northern; D003001:Cloning, Molecular; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism*; D016373:DNA, Antisense / Q000494:pharmacology; D054768:G-Protein-Coupled Receptor Kinases; D019204:GTP-Binding Proteins / Q000378:metabolism; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D011970:Receptors, Histamine H2 / Q000378:metabolism*; D013274:Stomach Neoplasms / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000378:metabolism; D051552:beta-Adrenergic Receptor Kinases
D016373:DNA, Antisense; D016376:Oligonucleotides, Antisense; D011970:Receptors, Histamine H2; D020794:Receptor Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; D017868:Cyclic AMP-Dependent Protein Kinases; D051552:beta-Adrenergic Receptor Kinases; D054768:G-Protein-Coupled Receptor Kinases; C089176:G-protein-coupled receptor kinase 6; D019204:GTP-Binding Proteins