Purification and characterization of a multifunctional calmodulin-dependent protein kinase from canine myocardial cytosol.

Purification and characterization of a multifunctional calmodulin-dependent protein kinase from canine myocardial cytosol.
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从犬心肌细胞质中纯化和表征多功能钙调蛋白依赖性蛋白激酶。

DOI:
10.1016/0003-9861(86)90396-6
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发表时间:
1986
影响因子:
3.9
通讯作者:
E. Miyamoto
E. Miyamoto
中科院分区:
生物学3区
文献类型:
--
作者:
T. Iwasa;N. Inoue;Kohji Fukunaga;Toshiaki Isobe;Tsuneo Okuyama;E. Miyamoto

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从犬心肌细胞质中提取钙调素依赖性蛋白激酶,纯化1150倍至表观均一性,产率为1.5%。纯化的酶具有aMr 550,000,沉降系数为16.6 S,并且通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测定,显示单一蛋白带,aMr 55,000(55 K蛋白)。纯化后的酶比活力为1.6 μmol/mg protein/min,以肌球蛋白轻链为底物,对钙调素和Ca ~(2+)的Ka值分别为67 nM和1.1 μM。钙调素与55 K蛋白结合。纯化的酶具有广泛的底物特异性。内源性蛋白质,包括糖原合酶,受磷蛋白,肌钙蛋白I从犬心脏磷酸化的酶。这些结果表明,纯化的酶作为一个多功能的蛋白激酶的Ca 2+,钙调素依赖的细胞功能的犬心肌,和酶类似的酶中检测到的大脑,肝脏和骨骼肌。
A calmodulin-dependent protein kinase from canine myocardial cytosol was purified 1150-fold to apparent homogeneity with a 1.5% yield. The purified enzyme had aMrof 550,000 with a sedimentation coefficient of 16.6 S, and showed a single protein band with aMrof 55,000 (55K protein), determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified enzyme had a specific activity of 1.6 μmol/mg protein/min, andKavalues of 67 nM and 1.1 μM for calmodulin and Ca2+, respectively, using chicken gizzard myosin light chain as substrate. Calmodulin bound to the 55K protein. The purified enzyme had a broad substrate specificity. Endogenous proteins including glycogen synthase, phospholamban, and troponin I from the canine heart were phosphorylated by the enzyme. These results suggest that the purified enzyme works as a multifunctional protein kinase in the Ca2+, calmodulin-dependent cellular functions of the canine myocardium, and that the enzyme resembles enzymes detected in the brain, liver, and skeletal muscle.
肌钙蛋白 I 磷酸化对牛心肌肌钙蛋白 Ca2 调节位点的 Ca2 结合特性的影响。
DOI: --
发表时间: 1982
期刊: The Journal of biological chemistry
影响因子: --
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DOI: --
发表时间: 1983
期刊: The Journal of biological chemistry
影响因子: --
作者:
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离体心脏肌浆网中钙调蛋白介导的钙转运调节和 (Ca2 Mg2 ) 激活的 ATP 酶活性。
DOI: --
发表时间: 1982
期刊: The Journal of biological chemistry
影响因子: --
作者:
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通讯作者: Antonetz,T