Isolation and Properties of YCK2, a Saccharomyces cerevisiae Homolog of Casein Kinase-1

• Published in: Archives of biochemistry and biophysics Vol. 305; no. 1; pp. 47 - 53
• Main Authors: Vancura, A., Oconnor, A., Patterson, S.D., Mirza, U., Chait, B.T., Kuret, J.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.08.1993
• Subjects: ACTIVIDAD ENZIMATICA; ACTIVITE ENZYMATIQUE; Amino Acid Sequence; Benzimidazoles - pharmacology; Casein Kinase I; Casein Kinases; CASEINA; CASEINE; Chemical Phenomena; Chemistry, Physical; Enzyme Stability; Guanosine Triphosphate - metabolism; Heparin - pharmacology; ISOENZIMAS; ISOENZYME; Isoquinolines - pharmacology; Kinetics; Molecular Sequence Data; PROPIEDADES FISICO-QUIMICAS; PROPRIETE PHYSICOCHIMIQUE; Protein Kinases - chemistry; Protein Kinases - isolation & purification; Protein Kinases - metabolism; PROTEINA QUINASA; PROTEINE KINASE; PURIFICACION; PURIFICATION; SACCHAROMYCES CEREVISIAE; Saccharomyces cerevisiae - enzymology; Saccharomyces cerevisiae Proteins; Substrate Specificity
• ISSN: 0003-9861; 1096-0384
• DOI: 10.1006/abbi.1993.1391

A soluble fragment of YCK2, a casein kinase-1 isoform from Saccharomyces cerevisiae, has been purified and characterized in vitro. The procedure enriches enzyme activity to a final specific activity of 4.7 μmol min −1 mg −1 (when assayed with casein as substrate). Structural analysis reveals that the preparation arises from N-terminal modification and C-terminal proteolysis of the initially synthesized 546-residue protein, consisting of residues 2-495 ± 1. Kinetic analysis demonstrates that YCK2 is similar to casein kinase-1 isolated from other organisms in its inability to use GTP as nucleotide substrate, in its sensitivity to heparin and ribofuranosylbenzimidazole inhibitors, and in its peptide substrate selectivity. The enzyme is unusual, however, in that it is insensitive to the potent mammalian casein kinase-1 inhibitor N-(2-aminoethyl)-5-chloroisoquinoline-8-sulfonamide.