{"title":"犬心脏部分纯化Na+,K+- atp酶的鉴定。","authors":"P V Sulakhe, V Elimban, N S Dhalla","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>Ouabain-sensitive Na+,K+-ATPase of isolated membranes represents a biochemical correlate of the \"Na+ pump\" that is present in intact tissue and is responsible for dissimilar distributions of Na+ and K+ across cellular plasma membranes. The enzyme has been purified from a variety of sources and its properties have been reported. Only a limited number of studies, however, deal with the cardiac Na+,K+-ATPase. We solubilized this enzyme from dog heart with deoxycholate and effected its further purification by NaI treatment. The method yielded an enzyme preparation of high specific activity (140 mumole/mg protein per hr). The following characteristics were noted: (1) pH optima of 7.4 and greater than 9.0 for ouabain-sensitive and -insensitive ATPases; (2) inhibition by Ca2+ and ouabain, the latter effect being allosteric in nature; (3) inhibition by sulfhydryl reagents (N-ethylmaleimide, p-chloromercuribenzoate) of the ouabain-sensitive ATPase but not of the -insensitive enzyme activity. All these properties resembled those seen in isolated plasma membranes (sarcolemma), suggesting that the purification procedure did not alter the properties of mono- and divalent interacting sites as well as a digitalis recognition domain of the Na+ pump. These results thus aid in further understanding the regulation of this vectorial pump that is critical in myocardial function.</p>","PeriodicalId":77831,"journal":{"name":"Advances in myocardiology","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1985-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Characterization of a partially purified Na+,K+-ATPase from dog heart.\",\"authors\":\"P V Sulakhe, V Elimban, N S Dhalla\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Ouabain-sensitive Na+,K+-ATPase of isolated membranes represents a biochemical correlate of the \\\"Na+ pump\\\" that is present in intact tissue and is responsible for dissimilar distributions of Na+ and K+ across cellular plasma membranes. The enzyme has been purified from a variety of sources and its properties have been reported. Only a limited number of studies, however, deal with the cardiac Na+,K+-ATPase. We solubilized this enzyme from dog heart with deoxycholate and effected its further purification by NaI treatment. The method yielded an enzyme preparation of high specific activity (140 mumole/mg protein per hr). The following characteristics were noted: (1) pH optima of 7.4 and greater than 9.0 for ouabain-sensitive and -insensitive ATPases; (2) inhibition by Ca2+ and ouabain, the latter effect being allosteric in nature; (3) inhibition by sulfhydryl reagents (N-ethylmaleimide, p-chloromercuribenzoate) of the ouabain-sensitive ATPase but not of the -insensitive enzyme activity. All these properties resembled those seen in isolated plasma membranes (sarcolemma), suggesting that the purification procedure did not alter the properties of mono- and divalent interacting sites as well as a digitalis recognition domain of the Na+ pump. These results thus aid in further understanding the regulation of this vectorial pump that is critical in myocardial function.</p>\",\"PeriodicalId\":77831,\"journal\":{\"name\":\"Advances in myocardiology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1985-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Advances in myocardiology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Advances in myocardiology","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Characterization of a partially purified Na+,K+-ATPase from dog heart.
Ouabain-sensitive Na+,K+-ATPase of isolated membranes represents a biochemical correlate of the "Na+ pump" that is present in intact tissue and is responsible for dissimilar distributions of Na+ and K+ across cellular plasma membranes. The enzyme has been purified from a variety of sources and its properties have been reported. Only a limited number of studies, however, deal with the cardiac Na+,K+-ATPase. We solubilized this enzyme from dog heart with deoxycholate and effected its further purification by NaI treatment. The method yielded an enzyme preparation of high specific activity (140 mumole/mg protein per hr). The following characteristics were noted: (1) pH optima of 7.4 and greater than 9.0 for ouabain-sensitive and -insensitive ATPases; (2) inhibition by Ca2+ and ouabain, the latter effect being allosteric in nature; (3) inhibition by sulfhydryl reagents (N-ethylmaleimide, p-chloromercuribenzoate) of the ouabain-sensitive ATPase but not of the -insensitive enzyme activity. All these properties resembled those seen in isolated plasma membranes (sarcolemma), suggesting that the purification procedure did not alter the properties of mono- and divalent interacting sites as well as a digitalis recognition domain of the Na+ pump. These results thus aid in further understanding the regulation of this vectorial pump that is critical in myocardial function.