{"title":"心脏组织中高活性瓦阿因敏感Na+, K+依赖性腺苷三磷酸酶的纯化和性质","authors":"Hideo Matsui, Arnold Schwartz","doi":"10.1016/0926-6593(66)90185-8","DOIUrl":null,"url":null,"abstract":"<div><p>A highly active and specific Na<sup>+</sup>, K<sup>+</sup>-dependent ATPase was obtained from calf-heart muscle by succesive treatments with deoxycholate and NaI.</p><p>General properties of the enzyme were studied. Simultaneous addition of Na<sup>+</sup> and K<sup>+</sup> increased the activity about 20 times above the basic Mg<sup>2+</sup>-dependent level. The activation was completely reversed by 10<sup>−4</sup> M ouabain. The optimal pH was 7.2; the <em>K</em><sub><em>m</em></sub> for ATP for the Na<sup>+</sup>,K<sup>+</sup>-dependent and Mg<sup>2+</sup>-dependent enzyme activities were 2.4·10<sup>−4</sup> M and 5.0·10<sup>−5</sup> M, respectively. Only CTP could be substituted for ATP, but the activity was 14% of that found with ATP.</p><p>Azide and histone, which affected the crude ATPase system, had no effect on the purified enzyme. <em>p</em>-Chloromercuribenzoate, <em>N</em>-ethylmaleimide, oligomycin, tributyltin chloride, octylguanidine as well as ouabain were found to inhibit the Na<sup>+</sup>, K<sup>+</sup>-dependent component while having little or no effect on the basic Mg<sup>2+</sup>-dependent activity. The <em>K</em><sub><em>i</em></sub>'s were 5·10<sup>−6</sup> M, 6·10<sup>−4</sup> M, 7·10<sup>−6</sup> M, 1.5·10<sup>−5</sup> M, 3·10<sup>−4</sup> M and 10<sup>−6</sup> M, respectively.</p></div>","PeriodicalId":100160,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Enzymology and Biological Oxidation","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1966-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0926-6593(66)90185-8","citationCount":"213","resultStr":"{\"title\":\"Purification and properties of a highly active ouabain-sensitive Na+, K+-dependent adenosinetriphosphatase from cardiac tissue\",\"authors\":\"Hideo Matsui, Arnold Schwartz\",\"doi\":\"10.1016/0926-6593(66)90185-8\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>A highly active and specific Na<sup>+</sup>, K<sup>+</sup>-dependent ATPase was obtained from calf-heart muscle by succesive treatments with deoxycholate and NaI.</p><p>General properties of the enzyme were studied. Simultaneous addition of Na<sup>+</sup> and K<sup>+</sup> increased the activity about 20 times above the basic Mg<sup>2+</sup>-dependent level. The activation was completely reversed by 10<sup>−4</sup> M ouabain. The optimal pH was 7.2; the <em>K</em><sub><em>m</em></sub> for ATP for the Na<sup>+</sup>,K<sup>+</sup>-dependent and Mg<sup>2+</sup>-dependent enzyme activities were 2.4·10<sup>−4</sup> M and 5.0·10<sup>−5</sup> M, respectively. Only CTP could be substituted for ATP, but the activity was 14% of that found with ATP.</p><p>Azide and histone, which affected the crude ATPase system, had no effect on the purified enzyme. <em>p</em>-Chloromercuribenzoate, <em>N</em>-ethylmaleimide, oligomycin, tributyltin chloride, octylguanidine as well as ouabain were found to inhibit the Na<sup>+</sup>, K<sup>+</sup>-dependent component while having little or no effect on the basic Mg<sup>2+</sup>-dependent activity. The <em>K</em><sub><em>i</em></sub>'s were 5·10<sup>−6</sup> M, 6·10<sup>−4</sup> M, 7·10<sup>−6</sup> M, 1.5·10<sup>−5</sup> M, 3·10<sup>−4</sup> M and 10<sup>−6</sup> M, respectively.</p></div>\",\"PeriodicalId\":100160,\"journal\":{\"name\":\"Biochimica et Biophysica Acta (BBA) - Enzymology and Biological Oxidation\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1966-11-15\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/0926-6593(66)90185-8\",\"citationCount\":\"213\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biochimica et Biophysica Acta (BBA) - Enzymology and Biological Oxidation\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/0926659366901858\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochimica et Biophysica Acta (BBA) - Enzymology and Biological Oxidation","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/0926659366901858","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Purification and properties of a highly active ouabain-sensitive Na+, K+-dependent adenosinetriphosphatase from cardiac tissue
A highly active and specific Na+, K+-dependent ATPase was obtained from calf-heart muscle by succesive treatments with deoxycholate and NaI.
General properties of the enzyme were studied. Simultaneous addition of Na+ and K+ increased the activity about 20 times above the basic Mg2+-dependent level. The activation was completely reversed by 10−4 M ouabain. The optimal pH was 7.2; the Km for ATP for the Na+,K+-dependent and Mg2+-dependent enzyme activities were 2.4·10−4 M and 5.0·10−5 M, respectively. Only CTP could be substituted for ATP, but the activity was 14% of that found with ATP.
Azide and histone, which affected the crude ATPase system, had no effect on the purified enzyme. p-Chloromercuribenzoate, N-ethylmaleimide, oligomycin, tributyltin chloride, octylguanidine as well as ouabain were found to inhibit the Na+, K+-dependent component while having little or no effect on the basic Mg2+-dependent activity. The Ki's were 5·10−6 M, 6·10−4 M, 7·10−6 M, 1.5·10−5 M, 3·10−4 M and 10−6 M, respectively.