Vancura A, O'Connor A, Patterson S D, Mirza U, Chait B T, Kuret J
Cold Spring Harbor Laboratory, New York 11724-2220.
Arch Biochem Biophys. 1993 Aug 15;305(1):47-53. 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 mumol 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 ribofuranosyl-benzimidazole 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.
来自酿酒酵母的酪蛋白激酶-1亚型YCK2的一种可溶性片段已在体外进行了纯化和特性鉴定。该方法将酶活性富集至最终比活性为4.7 μmol min⁻¹ mg⁻¹（以酪蛋白为底物进行测定时）。结构分析表明，该制剂源自最初合成的546个残基蛋白质的N端修饰和C端蛋白水解，由2 - 495 ± 1个残基组成。动力学分析表明，YCK2在不能将GTP用作核苷酸底物、对肝素和呋喃核糖基苯并咪唑抑制剂敏感以及肽底物选择性方面，与从其他生物体分离的酪蛋白激酶-1相似。然而，该酶不同寻常之处在于它对强效的哺乳动物酪蛋白激酶-1抑制剂N-(2-氨基乙基)-5-氯异喹啉-8-磺酰胺不敏感。
