Isolation and properties of YCK2, a Saccharomyces cerevisiae homolog of casein kinase-1.
Isolation and properties of YCK2, a Saccharomyces cerevisiae homolog of casein kinase-1.
复制标题
YCK2（一种酪蛋白激酶 1 的酿酒酵母同源物）的分离和特性。
DOI：
10.1006/abbi.1993.1391
复制
发表时间：
1993
影响因子：
3.9
通讯作者：
Kuret,J
中科院分区：
文献类型：
--
作者：
Vancura,A;O'Connor,A;Patterson,SD;Mirza,U;Chait,BT;Kuret,J
从酿酒酵母（Saccharomycescerevisiae）中分离得到酪蛋白激酶1（caseinkinase-1）的一个可溶性片段YCK 2。该方法将酶活性富集至最终比活性为4.7 μmol min− 1 mg −1（以酪蛋白为底物测定时）。结构分析表明，该制备物是由最初合成的546个残基蛋白（由残基2-495 ± 1组成）的N-末端修饰和C-末端蛋白水解产生的。动力学分析表明，YCK 2是类似的酪蛋白激酶-1分离自其他生物体在其无法使用GTP作为核苷酸底物，在其敏感性肝素和呋喃核糖基苯并咪唑抑制剂，并在其肽底物的选择性。然而，这种酶是不寻常的，因为它对有效的哺乳动物酪蛋白激酶-1激动剂N-（2-氨乙基）-5-氯异喹啉-8-磺酰胺不敏感。
A soluble fragment of YCK2, a casein kinase-1 isoform fromSaccharomyces cerevisiae, has been purified and characterizedin vitro. The procedure enriches enzyme activity to a final specific activity of 4.7 μmol min−1mg−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 inhibitorN-(2-aminoethyl)-5-chloroisoquinoline-8-sulfonamide.