The distribution and dissociation of cyclic adenosine 3':5'-monophosphate-dependent protein kinases in adipose, cardiac, and other tissues.
The distribution and dissociation of cyclic adenosine 3':5'-monophosphate-dependent protein kinases in adipose, cardiac, and other tissues.
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环腺苷 3:5-单磷酸依赖性蛋白激酶在脂肪、心脏和其他组织中的分布和解离。
DOI：
10.1016/s0021-9258(19)42003-6
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发表时间：
1975
期刊：
影响因子：
--
通讯作者：
C. R. Park
中科院分区：
文献类型：
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作者：
J. Corbin;S. L. Keely;C. R. Park
在脂肪组织的粗提物中，在 700 微克/毫升组蛋白或 0.5 M 氯化钠存在的情况下，蛋白激酶在 30 度时缓慢解离成调节和催化亚基。如果通过添加 10 muM 腺苷 3':5'-单磷酸 (cAMP) 首先解离激酶，则在通过 Sephadex G-25 层析去除 cAMP 后立即发生重新解离。相比之下，在心脏粗提取物中，蛋白激酶在 700 微克/毫升组蛋白或 0.5 M 氯化钠存在下快速解离，并在去除 cAMP 后缓慢重新结合。这些差异是由于这些组织中存在两种类型的蛋白激酶（称为 I 型和 II 型）造成的。脂肪组织提取物的 DEAE-纤维素层析仅产生 cAMP 依赖性蛋白激酶活性（II 型）的一个峰，该峰在 0.15 至 0.25 M NaCl 之间洗脱。心脏提取物的类似色谱法将酶活性分解为两个峰； I 型酶在 0.05 至 0.1 M 之间洗脱并占主导地位（大于总活性的 75%），以及在 0.15 至 0.25 M NaCl 之间洗脱的 II 型酶。通过 DEAE-纤维素和 Sepharose 6B 层析部分纯化后，心脏和脂肪组织中的 I 型和 II 型酶的解离特性得以保留。对分离的心肌酶峰进行重色谱分析不会改变洗脱模式。蔗糖密度梯度离心和凝胶过滤研究表明这些酶的分子量非常相似。通过心脏提取物的 DEAE-纤维素色谱分离的 II 型酶类似于脂肪组织酶，即在组蛋白或 0.5 M NaCl 存在的情况下，它在 30 度下缓慢解离。然而，脂肪组织激酶和心脏 II 型激酶并不相同，因为它们在 DEAE-纤维素柱上的洗脱点并不完全相同。对几种组织的调查表明，存在与脂肪组织和心脏中的酶相似的 I 型和 II 型蛋白激酶，通过粗提物的 DEAE-纤维素色谱法以及通过酶与组蛋白的解离来确定。 MgATP 的存在可防止 0.5 M 氯化钠或组蛋白使 I 型酶从心脏解离。然而，DEAE-纤维素上酶的特性并没有改变......
In crude extracts of adipose tissue the protein kinase dissociates slowly at 30 degrees into regulatory and catalytic subunits in the presence of 700 mug per ml of histone or 0.5 M NaCl. If the kinase is first dissociated by adding 10 muM adenosine 3':5'-monophosphate (cAMP), reassociation occurs instantaneously after removal of the cAMP by Sephadex G-25 chromatography. In contrast, in crude xtracts of heart, the protein kinase dissociates rapidly in the presence of 700 mug per ml of histone or 0.5 M NaCl and reassociates slowly after removal of cAMP. These differences are accounted for by the existence of two types of protein kinases in these tissues, referred to as types I and II. DEAE-cellulose chromatography of extracts of adipose tissue produces only one peak of cAMP-dependent protein kinase activity (type II) which elutes between 0.15 and 0.25 M NaCl. Similar chromatography of heart extracts resolves enzyme activity into two peaks; a type I enzyme which elutes between 0.05 and 0.1 M and predominates (greater than 75% of total activity), and a type II enzyme which elutes between 0.15 and 0.25 M NaCl. The dissociation properties of the types I and II enzymes from heart and adipose tissue are retained after partial purification by DEAE-cellulose and Sepharose 6B chromatography. Rechromatography of the separated peaks of the cardiac enzymes does not change the elution pattern. Sucrose density gradient centrifugation and gel filtration studies indicate that the molecular weights of these enzymes are very similar. The type II enzyme isolated by DEAE-cellulose chromatography of heart extracts resembles the adipose tissue enzyme, i.e. it undergoes slow dissociation at 30 degrees in the presence of histone or 0.5 M NaCl. The adipose tissue kinase and the heart type II kinase are not identical, however, since they do not elute at exactly the same point on DEAE-cellulose columns. A survey of several tissues indicates the presence of type I and II protein kinases similar to the enzymes in adipose tissue and heart as determined by DEAE-cellulose chromatography of crude extracts and by dissociation of the enzymes with histone. The presence of MgATP prevents dissociation of type I enzyme from heart by 0.5 M NaCl or histone. The profile of the enzyme on DEAE-cellulose, however, is not changed...