Detection of cGMP dependent protein kinase isozymes by specific antibodies.
Two isozymes of cGMP-dependent protein kinase (cGMP kinase) have been identified. Polyclonal antibodies were developed which recognize both isozymes or specifically the I alpha and I beta isoform. The specificity of these antibodies was verified by using the recombinant or purified I alpha and I beta isozymes. The antibodies cross-reacted with the purified isozymes of cGMP kinase from bovine tracheal smooth muscle. The tissue concentration of cGMP kinase was determined by ELISA. High concentrations (greater than 10 pmol/g wet tissue) were present in bovine lung, rumen, trachea, aorta, uterus and stomach. The tissue distribution of the isozymes I alpha and I beta was investigated by immunoblots using crude extracts of the different tissues. The I beta-specific antibody yielded strong signals with extracts of trachea, aorta, stomach and uterus, whereas heart, cerebellum and lung apparently contain mainly the I alpha isozyme.
Keilbach
A
A
Institut für Pharmakologie und Toxikologie, Technische Universität München, Federal Republic of Germany.
Ruth
P
P
Hofmann
F
F
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D016466:CHO Cells; D002417:Cattle; D006224:Cricetinae; D006152:Cyclic GMP / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015151:Immunoblotting; D007527:Isoenzymes / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D008969:Molecular Sequence Data; D009928:Organ Specificity; D011494:Protein Kinases / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D014018:Tissue Distribution
D000906:Antibodies; D007527:Isoenzymes; D011494:Protein Kinases; D006152:Cyclic GMP