A newly synthesized selective casein kinase I inhibitor, N-(2-aminoethyl)-5-chloroisoquinoline-8-sulfonamide, and affinity purification of casein kinase I from bovine testis.
When screening various isoquinolinesulfonamide compounds which we synthesized, CKI-7, N-(2-amino-ethyl)-5-chloroisoquinoline-8-sulfonamide, was found to have a potent inhibitory action against casein kinase I and a much weaker effect on casein kinase II and other protein kinases. Kinetic analysis indicated that CKI-7 inhibited casein kinase I competitively with respect to ATP and that the Ki values were 8.5 microM for casein kinase I and 70 microM for casein kinase II. An affinity chromatography absorbent was synthesized by coupling CKI-8 (1-(5-chloroisoquinoline-8-sulfonyl], a derivative of CKI-7, to cyanogen bromide-activated Sepharose 4B. Partially purified casein kinase I from bovine testis was subjected to affinity chromatography. Analysis of the purified casein kinase I by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate revealed a single band with molecular weight 37,000. These newly synthesized compounds, CKI-7 and CKI-8, should serve as useful tools for elucidating the biological significance of casein kinase I-mediated reactions.
Chijiwa
T
T
Department of Pharmacology, Nagoya University School of Medicine, Japan.
Hagiwara
M
M
Hidaka
H
H
D000818:Animals; D047388:Casein Kinases; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002846:Chromatography, Affinity; D007546:Isoquinolines / Q000494:pharmacology*; D007700:Kinetics; D008297:Male; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000302:isolation & purification; D011817:Rabbits; D013379:Substrate Specificity; D013737:Testis / Q000201:enzymology*
D007546:Isoquinolines; D047428:Protein Kinase Inhibitors; C058815:N-(2-aminoethyl)-5-chloroisoquinoline-8-sulfonamide; D011494:Protein Kinases; D047388:Casein Kinases; D011493:Protein Kinase C