Casein Kinase II-Type Protein Kinase from Pea Cytoplasm and Its Inactivation by Alkaline Phosphatase in Vitro
Casein Kinase II-Type Protein Kinase from Pea Cytoplasm and Its Inactivation by Alkaline Phosphatase in Vitro
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豌豆细胞质酪蛋白激酶II型蛋白激酶及其体外碱性磷酸酶灭活作用
DOI：
10.1104/pp.103.3.955
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
S. Roux
中科院分区：
文献类型：
--
作者：
S. Zhang;C. Jin;S. Roux
从黄化豌豆（PisumsativumL.）根据考马斯蓝染色的十二烷基硫酸钠-聚丙烯酰胺凝胶判断，胚芽纯度约为90%。该激酶具有四聚体[alpha][alpha]′[beta]2结构，天然分子量为150 kD，两个催化亚基（[alpha]和[alpha]′）的亚基分子量为41和40 kD，推定的调节亚基（[beta]）的亚基分子量为35 kD。丝氨酸和苏氨酸残基磷酸化时，混合酪蛋白，[β]-酪蛋白，或卵黄蛋白作为底物，而只有丝氨酸磷酸化，如果[α]-酪蛋白或组蛋白III-S是基板。0.5 mM精胺（50%最大酶活性所需的浓度[A50] = 0.1 mM）和2.5 mM亚精胺（A50 = 0.4 mM）的激酶活性刺激130%和80%，而腐胺和尸胺没有影响。激酶对肝素抑制非常敏感（50%抑制浓度[I50] = 0.025 [mu]g/mL）。与大多数其他酪蛋白激酶II型蛋白激酶相反，该制剂被K+和Na+抑制，I50值分别为75和65 mM。在体外用碱性磷酸酶预处理纯化的激酶制剂，导致其活性降低5倍。此外，当它的β亚基在没有底物的情况下自磷酸化时，这种激酶也失去了活性。这些结果表明，这种酪蛋白激酶II蛋白激酶的活性可能受到其多聚体结构中两个不同位点的磷酸化状态的调节。
A casein kinase II-type protein kinase has been purified from the cytosolic fraction of etiolated pea (Pisum sativum L.) plumules to about 90% purity as judged from Coomassie blue stained sodium dodecyl sulfate-polyacrylamide gels. This kinase has a tetrameric [alpha][alpha]′[beta]2 structure with a native molecular mass of 150 kD, and subunit molecular masses of 41 and 40 kD for the two catalytic subunits ([alpha] and [alpha]′) and 35 kD for the putative regulatory subunit ([beta]).Casein and phosvitin can be used as artificial substrates for this kinase. Both serine and threonine residues were phosphorylated when mixed casein, [beta]-casein, or phosvitin were used as the substrate, whereas only serine was phosphorylated if [alpha]-casein or histone III-S was the substrate. The kinase activity was stimulated 130% by 0.5 mM spermine (the concentration required for 50% of maximal enzyme activity [A50] = 0.1 mM) and 80% by 2.5 mM spermidine (A50 = 0.4 mM), whereas putrescine and cadaverine had no effect. The kinase was very sensitive to inhibition by heparin (concentration for 50% inhibition [I50] = 0.025 [mu]g/mL). In contrast to most other casein kinase II-type protein kinases, this preparation was inhibited by K+ and Na+, with I50 values of 75 and 65 mM, respectively. Pretreatment of the purified kinase preparation in vitro with alkaline phosphatase caused a 5-fold decrease in its activity. Additionally, this kinase also lost its activity when its [beta] subunit was autophosphorylated in the absence of substrate. These results suggest that the activity of this casein kinase II protein kinase may be regulated by the phosphorylation state of two different sites in its multimeric structure.
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DOI：
--
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Yan,TF;Tao,M
通讯作者：
Tao,M
DOI：
10.1042/bj2830021
发表时间：
1992-04
期刊：
The Biochemical journal
影响因子：
--
作者：
D. Sacks;H. Davis;J. Williams;E. Sheehan;J. Garcia;J. McDonald
通讯作者：
D. Sacks;H. Davis;J. Williams;E. Sheehan;J. Garcia;J. McDonald
酪蛋白激酶——多能蛋白激酶。
DOI：
--
发表时间：
1982
期刊：
Current topics in cellular regulation
影响因子：
--
作者：
Hathaway,GM;Traugh,JA
通讯作者：
Traugh,JA
西兰花酪蛋白激酶 II 的磷酸化可刺激拟南芥 G-box 结合因子 GBF1 的 DNA 结合活性。
DOI：
10.1105/tpc.4.1.87
发表时间：
1992
期刊：
The Plant cell
影响因子：
--
作者：
Klimczak,LJ;Schindler,U;Cashmore,AR
通讯作者：
Cashmore,AR
影响因子：
11.6
作者：
DATTA, N;CASHMORE, AR
通讯作者：
CASHMORE, AR