Stimulation of the thiol-dependent ADP-ribosyltransferase and NAD glycohydrolase activities of Bordetella pertussis toxin by adenine nucleotides, phospholipids, and detergents.
Stimulation of the thiol-dependent ADP-ribosyltransferase and NAD glycohydrolase activities of Bordetella pertussis toxin by adenine nucleotides, phospholipids, and detergents.
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腺嘌呤核苷酸、磷脂和去污剂刺激百日咳博德特氏菌毒素的硫醇依赖性 ADP-核糖基转移酶和 NAD 糖水解酶活性。
DOI：
10.1021/bi00357a066
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发表时间：
1986
期刊：
影响因子：
2.9
通讯作者：
Hewlett,EL
中科院分区：
文献类型：
--
作者：
Moss,J;Stanley,SJ;Watkins,PA;Burns,DL;Manclark,CR;Kaslow,HR;Hewlett,EL
弗吉尼亚大学医学院医学和药理学系，Charlottesville，Virginia 22908接收1985年10月23日摘要：百日咳毒素催化鸟苷酸结合蛋白转导素的ADP-核糖基化被腺嘌呤核苷酸和磷脂或去污剂刺激。为了确定这些试剂的作用位点，检查了它们对非转导蛋白依赖性NAD糖水解酶活性的影响。毒素催化的NAD水解通过ATP和洗涤剂或磷脂协同增加;两性离子洗涤剂3-[（3-胆酰胺丙基）二甲基铵基]-1-丙磺酸盐（CHAPS）比非离子洗涤剂Triton X-100>溶血磷脂酰胆碱>磷脂酰胆碱更有效。在CHAPS存在下，ATP的Ao s为2.6| tM;在胆酸盐或溶血磷脂酰胆碱存在下，最大激活需要显著更高浓度的ATP。在CHAPS中，NAD的水解被ATP> ADP> AMP>腺苷增强; ATP比MgATP或不可水解的类似物腺苷-5'-基亚氨基二磷酸更有效。GTP和脒基-5'-基亚氨基二磷酸的活性低于相应的腺嘌呤核苷酸。在CHAPS和ATP的存在下的活性几乎完全依赖于二硫苏糖醇;二硫苏糖醇的Ao s被CHAPS单独显着降低，并在更大程度上，由CHAPS和ATP。为了确定ATP、CHAPS和二硫苏糖醇的作用位点，通过色谱法分离酶（Sj）和结合组分（B寡聚体）。纯化的Si亚基催化NAD的二硫苏糖醇依赖性水解;活性通过CHAPS而不是ATP增强。这些研究与腺嘌呤核苷酸、二硫苏糖醇和CHAPS作用于毒素本身而不是作用于底物的结论是一致的;腺嘌呤核苷酸似乎参与毒素的活化而不是分离的催化单元。
Departments of Medicine and Pharmacology, University of Virginia School of Medicine, Charlottesville, Virginia 22908 Received October 23, 1985 abstract: Pertussis toxin catalyzed ADP-ribosylation of the guanyl nucleotide binding protein transducin was stimulated by adenine nucleotide and either phospholipids or detergents. To determine the sites of action of these agents, their effects were examined on the transducin-independent NAD glycohydrolase activity. Toxin-catalyzed NAD hydrolysis was increased synergistically by ATP and detergents or phospholipids; the zwitterionic detergent 3-[(3-cholamidopropyl) dimethylammonio]-l-propanesulfonate (CHAPS) was more effective than the nonionic detergent Triton X-100> lysophosphatidylcholine> phosphatidylcholine. The Ao s for ATP inthe presence of CHAPS was 2.6| tM; significantly higher concentrations of ATP were required for maximal activation in the presence of cholate or lysophosphatidylcholine. In CHAPS, NAD hydrolysis was enhanced by ATP> ADP> AMP> adenosine; ATP was more effective than MgATP or the nonhydrolyzable analogue adenyl-5'-yl imidodiphosphate. GTP and guanyl-5'-yl imidodiphosphate were less active than the corresponding adenine nucleotides. Activity in the presence of CHAPS and ATP was almost completely dependent on dithiothreitol; the Ao s for dithiothreitol was significantly decreased by CHAPS alone and, to a greater extent, by CHAPS and ATP. To determine the site of action of ATP, CHAPS, and dithiothreitol, the enzymatic (Sj) and binding components (B oligomer) were resolved by chromatography. The purified Si subunit catalyzed the dithiothreitol-dependent hydrolysis of NAD; activity was enhanced by CHAPS but not ATP. Thestudies are consistent with the conclusion that adenine nucleotides, dithiothreitol, and CHAPS act on the toxin itself rather than on the substrate; adenine nucleotides appear to be involved in the activation of toxin but not the isolated catalytic unit.