Inhibition of the adenosinetriphosphatase activity of Escherichia coli F1 by the water-soluble carbodiimide 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide is due to modification of several carboxyls in the beta subunit.
Inhibition of the adenosinetriphosphatase activity of Escherichia coli F1 by the water-soluble carbodiimide 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide is due to modification of several carboxyls in the beta subunit.
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水溶性碳二亚胺1-乙基-3-[3-(二甲基氨基)丙基]碳二亚胺对大肠杆菌F1的腺苷三磷酸酶活性的抑制是由于β亚基中几个羧基的修饰。
DOI：
10.1021/bi00313a019
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发表时间：
1984
期刊：
影响因子：
2.9
通讯作者：
Capaldi,RA
中科院分区：
文献类型：
--
作者：
Lötscher,HR;deJong,C;Capaldi,RA
摘要：L-乙基-3-[3-(二甲氨基)丙基]碳二亚胺(EDC)对大肠杆菌ATPase的活性有时间和浓度依赖性的抑制作用。Mg~(2+)离子能显著降低酶的失活。每摩尔ECF中近13摩尔的EDC被掺入酶中，对ATPase活性的抑制率为95%。三分之二的标记被发现与B亚基有关，其化学计量比约为每摩尔ft约3摩尔的EDC。用溴化氰切EDC修饰的B亚基，用高压液相色谱分离得到33个氨基酸(CB8，残基162-194)，每摩尔肽含有3摩尔的EDC。在胰酶多肽图谱中，可以识别出两个EDC标记片段(T18，残基166-183，和T20，残基186-202)。EDC诱导的B亚基内部显著的内部交联会使分析变得复杂。结果表明，EDC在B亚基的短片段上修饰了多个位点，其中包括二环己基碳二亚胺修饰的谷氨酸。
Hans-Ruedi Lotscher, Catherina deJong, and Roderick A. Capaldi* abstract: Reaction of the ATPase of Escherichia coli (ECF,) with l-ethyl-3-[3-(dimethylamino) propyl] carbodiimide (EDC) resulted in a time-and concentration-dependent in-hibition of ATPase activity. The inactivation was greatly reduced by Mg2+ ions. Close to 13 mol of EDC per mol of ECF, was incorporated into the enzyme at95% inhibition of ATPase activity. Two-thirdsof the label was found to be associated with subunit B with a stoichiometry of about 3 mol of EDC per mol of ft. Cleavage of EDC-modifiedsubunit B with cyanogen bromide and fractionation of the peptides by high-pressure liquid chromatography revealed a short segment of 33 amino acids (CB8, residues 162-194) containing 3 mol of EDC per mol of peptide. In tryptic peptide maps, two EDC-labeled fragments could be identified (T18, residues 166-183, and T20, residues 186-202). The analyses were complicated by significant internal cross-linking within the B subunit induced by EDC. The results show that EDC modifies multiple sites in a short segment of subunit B which includes the glutamic acids modified by dicyclohexylcarbodiimide in