{"title":"泌乳兔乳腺酪蛋白激酶II的纯化及特性研究","authors":"Vanio Mitev, Alain Pauloin, Louis-Marie Houdebine","doi":"10.1016/0020-711X(94)90167-8","DOIUrl":null,"url":null,"abstract":"<div><p></p><ul><li><span>1.</span><span><p>1. Highly purified 200 kDa casein kinase II from rabbit lactating mammary gland (MG-CK II) was obtained by means of a new purification procedure consisting of one phosphocellulose and three Mono Q steps.</p></span></li><li><span>2.</span><span><p>2. Its <em>K</em><sub><em>m</em></sub> for ATP was 2.22 μM and 0.57 mg/ml and 0.13 mg/ml for partially dephosphorylated casein and phosvitin respectively. Stathmine was also suitable as substrate. 2-aminopurine and 6-dimethylaminopurine inhibited efficiently MG-CK II <em>K</em><sub><em>i</em></sub> = 5 and 1 mM respectively).</p></span></li><li><span>3.</span><span><p>3. MG-CK II autophosphorylated on its α-, α '- and β-subunits. The β-subunit auto-phosporylation was enhanced in presence of exogenous substrate. Its modulation was highly dependent on ATP concentration.</p></span></li><li><span>4.</span><span><p>4. The effects of basic compounds which affected dramatically the phosphorylation of dephosphorylated casein in presence of various ATP concentrations were reported.</p></span></li></ul></div>","PeriodicalId":13733,"journal":{"name":"International Journal of Biochemistry","volume":"26 5","pages":"Pages 667-677"},"PeriodicalIF":0.0000,"publicationDate":"1994-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0020-711X(94)90167-8","citationCount":"3","resultStr":"{\"title\":\"Purification and characterization of casein kinase II from lactating rabbit mammary gland\",\"authors\":\"Vanio Mitev, Alain Pauloin, Louis-Marie Houdebine\",\"doi\":\"10.1016/0020-711X(94)90167-8\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p></p><ul><li><span>1.</span><span><p>1. Highly purified 200 kDa casein kinase II from rabbit lactating mammary gland (MG-CK II) was obtained by means of a new purification procedure consisting of one phosphocellulose and three Mono Q steps.</p></span></li><li><span>2.</span><span><p>2. Its <em>K</em><sub><em>m</em></sub> for ATP was 2.22 μM and 0.57 mg/ml and 0.13 mg/ml for partially dephosphorylated casein and phosvitin respectively. Stathmine was also suitable as substrate. 2-aminopurine and 6-dimethylaminopurine inhibited efficiently MG-CK II <em>K</em><sub><em>i</em></sub> = 5 and 1 mM respectively).</p></span></li><li><span>3.</span><span><p>3. MG-CK II autophosphorylated on its α-, α '- and β-subunits. The β-subunit auto-phosporylation was enhanced in presence of exogenous substrate. Its modulation was highly dependent on ATP concentration.</p></span></li><li><span>4.</span><span><p>4. The effects of basic compounds which affected dramatically the phosphorylation of dephosphorylated casein in presence of various ATP concentrations were reported.</p></span></li></ul></div>\",\"PeriodicalId\":13733,\"journal\":{\"name\":\"International Journal of Biochemistry\",\"volume\":\"26 5\",\"pages\":\"Pages 667-677\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1994-05-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/0020-711X(94)90167-8\",\"citationCount\":\"3\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"International Journal of Biochemistry\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/0020711X94901678\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"International Journal of Biochemistry","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/0020711X94901678","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 3
摘要
1.1. 采用一个磷酸纤维素和三个Mono Q步骤纯化兔泌乳乳腺酪蛋白激酶(MG-CK II)，获得了高纯度的200 kDa酪蛋白激酶。其对ATP的Km为2.22 μM，对部分去磷酸化酪蛋白和磷维素的Km分别为0.57 mg/ml和0.13 mg/ml。石竹碱也适合作为底物。2-氨基嘌呤和6-二甲氨基嘌呤有效抑制MG-CK II Ki = 5和1 mM)。MG-CK II在其α-、α '-和β-亚基上发生自磷酸化。外源底物存在时，β亚基自磷酸化增强。其调节高度依赖于ATP浓度。报道了碱性化合物在不同ATP浓度下对去磷酸化酪蛋白磷酸化的影响。
Purification and characterization of casein kinase II from lactating rabbit mammary gland
1.
1. Highly purified 200 kDa casein kinase II from rabbit lactating mammary gland (MG-CK II) was obtained by means of a new purification procedure consisting of one phosphocellulose and three Mono Q steps.
2.
2. Its Km for ATP was 2.22 μM and 0.57 mg/ml and 0.13 mg/ml for partially dephosphorylated casein and phosvitin respectively. Stathmine was also suitable as substrate. 2-aminopurine and 6-dimethylaminopurine inhibited efficiently MG-CK II Ki = 5 and 1 mM respectively).
3.
3. MG-CK II autophosphorylated on its α-, α '- and β-subunits. The β-subunit auto-phosporylation was enhanced in presence of exogenous substrate. Its modulation was highly dependent on ATP concentration.
4.
4. The effects of basic compounds which affected dramatically the phosphorylation of dephosphorylated casein in presence of various ATP concentrations were reported.
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