Inactivation of C30A trimethylamine dehydrogenase by N-cyclopropyl-alpha-methylbenzylamine, 1-phenylcyclopropylamine, and phenylhydrazine.
Inactivation of C30A trimethylamine dehydrogenase by N-cyclopropyl-alpha-methylbenzylamine, 1-phenylcyclopropylamine, and phenylhydrazine.
复制标题
N-环丙基-α-甲基苄胺、1-苯基环丙胺和苯肼使 C30A 三甲胺脱氢酶失活。
DOI：
10.1021/bi0105126
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发表时间：
2001
期刊：
影响因子：
2.9
通讯作者：
Silverman,RB
中科院分区：
文献类型：
--
作者：
Mitchell,DJ;Nikolic,D;Jang,MH;vanBreemen,RB;Hille,R;Silverman,RB
Trimethylamine dehydrogenase (TMADH) from the bacteriumMethylophilus methylotrophus(sp. W3A1) and its C30A mutant were inactivated with three known inactivators of monoamine oxidase, namely, phenylhydrazine,N-cyclopropyl-α-methylbenzylamine, and 1-phenylcyclopropylamine. All three compounds irreversibly inactivated both the wild-type and C30A mutant enzymes, although phenylhydrazine was 10 times more potent thanN-cyclopropyl-α-methylbenzylamine, which was much more potent than 1-phenylcyclopropylamine. The change in the UV−visible absorption spectra upon modification indicated that the flavin was modified. In the case of the C30A mutant, the absence of a covalent attachment of the flavin to the polypeptide has permitted LC-electrospray mass spectrometry of the reaction product to be undertaken, demonstrating new mass peaks corresponding to various chemically modified forms of the flavin cofactor. In the case ofN-cyclopropyl-α-methylbenzylamine, masses corresponding to hydroxy-FMN and hydroxyriboflavin were detected. 1-Phenylcyclopropylamine inactivation of the C30A mutant produced three modified flavins, as evidenced by the electrospray mass spectrum: hydroxy-FMN, FMN plus C6H5COCH2CH2, and hydroxy-FMN plus C6H5COCH2CH2. Phenylhydrazine inactivation of the C30A mutant gave at least seven different modified flavins: hydroxyriboflavin, hydroxy-FMN, two apparently isomeric compounds corresponding to hydroxy-FMN plus one phenyl group, two apparently isomeric compounds corresponding to FMN plus one phenyl group, and FMN plus two phenyl groups. Covalent flavin adduct formation appears to be the only modification because dialysis of the inactive enzyme followed by reconstitution with FMN restores the enzyme activity to that of a noninactivated control.
