{"title":"底物与丙酰辅酶a羧化酶结合的结构要求","authors":"C.S. Hegre, M.Daniel Lane","doi":"10.1016/0926-6593(66)90154-8","DOIUrl":null,"url":null,"abstract":"<div><p>Further purification of propionyl-CoA carboxylase (EC 6.4.1.3) from bovine-liver mitochondria is reported. On the basis of the present and earlier investigations it is concluded that binding of acyl-CoA substrates by the carboxylase occurs through the CoA moiety. The relative maximum velocities of propionyl-CoA, propionyl-dephospho-CoA, and propionyl-pantetheine carboxylation were 1.0, 0.29, and 0.014, respectively; the <span><math><mtext>K</mtext><msub><mi></mi><mn>m</mn></msub></math></span> values for the same substrates were 2.6·10<sup>−4</sup>, 2.8·10<sup>−3</sup>, and 2.4·10<sup>−2</sup> M, respectively. Enzymatic carboxylation of <span><math><mtext>propionyl</mtext><mtext>-N-</mtext><mtext>acetylcysteamine</mtext></math></span> could not be demonstrated. Coenzyme A and 3′-AMP were found to inhibit the carboxylation reaction competitively with respect to propionyl-CoA, whereas, inhibition by 2′-AMP and 5′-AMP was of a mixed type. The binding of acyl-CoA substrate to the carboxylase appears to involve the 3′-phosphate, adenine, and pantoyl moieties of the subsrate. Propionyl-CoA carboxylase is protected from <span><math><mtext>p-</mtext><mtext>chloromercuribenzoate</mtext></math></span> inhibition by propionyl-CoA, whereas, ATP-MgCl<sub>2</sub> facilitates this inhibition.</p></div>","PeriodicalId":100160,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Enzymology and Biological Oxidation","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1966-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0926-6593(66)90154-8","citationCount":"7","resultStr":"{\"title\":\"Structural requirements for substrate binding to propionyl-CoA carboxylase\",\"authors\":\"C.S. Hegre, M.Daniel Lane\",\"doi\":\"10.1016/0926-6593(66)90154-8\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Further purification of propionyl-CoA carboxylase (EC 6.4.1.3) from bovine-liver mitochondria is reported. On the basis of the present and earlier investigations it is concluded that binding of acyl-CoA substrates by the carboxylase occurs through the CoA moiety. The relative maximum velocities of propionyl-CoA, propionyl-dephospho-CoA, and propionyl-pantetheine carboxylation were 1.0, 0.29, and 0.014, respectively; the <span><math><mtext>K</mtext><msub><mi></mi><mn>m</mn></msub></math></span> values for the same substrates were 2.6·10<sup>−4</sup>, 2.8·10<sup>−3</sup>, and 2.4·10<sup>−2</sup> M, respectively. Enzymatic carboxylation of <span><math><mtext>propionyl</mtext><mtext>-N-</mtext><mtext>acetylcysteamine</mtext></math></span> could not be demonstrated. Coenzyme A and 3′-AMP were found to inhibit the carboxylation reaction competitively with respect to propionyl-CoA, whereas, inhibition by 2′-AMP and 5′-AMP was of a mixed type. The binding of acyl-CoA substrate to the carboxylase appears to involve the 3′-phosphate, adenine, and pantoyl moieties of the subsrate. Propionyl-CoA carboxylase is protected from <span><math><mtext>p-</mtext><mtext>chloromercuribenzoate</mtext></math></span> inhibition by propionyl-CoA, whereas, ATP-MgCl<sub>2</sub> facilitates this inhibition.</p></div>\",\"PeriodicalId\":100160,\"journal\":{\"name\":\"Biochimica et Biophysica Acta (BBA) - Enzymology and Biological Oxidation\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1966-10-17\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/0926-6593(66)90154-8\",\"citationCount\":\"7\",\"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/0926659366901548\",\"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/0926659366901548","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 7
摘要
进一步从牛肝线粒体中纯化丙酰辅酶a羧化酶(EC 6.4.1.3)。根据目前和早期的研究,可以得出结论,羧化酶通过辅酶a片段结合酰基辅酶a底物。丙酰辅酶a、丙酰去磷酸辅酶a和丙酰泛氨酸羧化反应的相对最大速度分别为1.0、0.29和0.014;相同基质的Km值分别为2.6·10−4、2.8·10−3和2.4·10−2 M。不能证明丙炔- n -乙酰半胱胺的酶羧化作用。辅酶A和3′-AMP对丙酰辅酶A的羧化反应具有竞争性抑制作用,而2′-AMP和5′-AMP的抑制作用为混合型。酰基辅酶a底物与羧化酶的结合似乎涉及底物的3 ' -磷酸、腺嘌呤和泛酰基部分。丙烯酰辅酶a羧化酶不受丙烯酰辅酶a对氯脲苯甲酸盐的抑制,而ATP-MgCl2促进了这种抑制。
Structural requirements for substrate binding to propionyl-CoA carboxylase
Further purification of propionyl-CoA carboxylase (EC 6.4.1.3) from bovine-liver mitochondria is reported. On the basis of the present and earlier investigations it is concluded that binding of acyl-CoA substrates by the carboxylase occurs through the CoA moiety. The relative maximum velocities of propionyl-CoA, propionyl-dephospho-CoA, and propionyl-pantetheine carboxylation were 1.0, 0.29, and 0.014, respectively; the values for the same substrates were 2.6·10−4, 2.8·10−3, and 2.4·10−2 M, respectively. Enzymatic carboxylation of could not be demonstrated. Coenzyme A and 3′-AMP were found to inhibit the carboxylation reaction competitively with respect to propionyl-CoA, whereas, inhibition by 2′-AMP and 5′-AMP was of a mixed type. The binding of acyl-CoA substrate to the carboxylase appears to involve the 3′-phosphate, adenine, and pantoyl moieties of the subsrate. Propionyl-CoA carboxylase is protected from inhibition by propionyl-CoA, whereas, ATP-MgCl2 facilitates this inhibition.