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首页> 外文期刊>Biochemistry >Spectrometic Evidence for the Flavin-1-Phenylcyclopropylamine Inactivator Adduct with Monoamine Oxidase N
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Spectrometic Evidence for the Flavin-1-Phenylcyclopropylamine Inactivator Adduct with Monoamine Oxidase N

机译：黄素-1-苯基环丙胺灭活剂与单胺氧化酶N的光谱学证据

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摘要

1-Phenylcyclopropylamine (1-PCPA) is shown to be an inactivator of the fungal flavoenzyme monoamine oxidase (MAO) N. Inactivation results in an increase in absorbance at 410 nm and is accompanied by the concomitant loss of the flavin absorption band at 458 nm. The spectral properties of the covalent adduct formed between the flavin cofactor of MAO N and I -PCPA are similar to those reported for the irreversible inactivation product formed with 1 -PCPA and mammalian mitochondrial monoamine oxidase B [Silverman, R. B., and Zieske, P. A. (1985) Biochemistry 24, 2128—2138]. There is a hypsochromic shift of the 410 nm band upon lowering the pH to 2, indicating that an N5-flavin adduct formed upon inactivation. Use of the fungal enzyme, MAO N, which lacks the covalent attachment to the flavin adenine dinucleotide (FAD) cofactor present in the mammalian forms MAO A and MAO B, has allowed for the isolation and further structural identification of the flavin—inactivator adduct. The incorporation of two 13C labels into the inactivator, [2,3-’3C2]-1-PCPA, followed by analysis using on-line liquid chromatography/electrospray ionization mass spectrometry and nuclear magnetic resonance spectroscopy, provided a means to explore the structure of the flavin—inactivator adduct of MAO N. The spectral evidence supports covalent attachment of the 1-PCPA inactivator to the cofactor as N5-3-oxo-3-phenylpropyl-FAD.
机译：1-苯基环丙胺（1-PCPA）被证明是真菌黄酮酶单胺氧化酶（MAO）N的失活剂。失活导致410 nm处吸光度增加，并伴随着458 nm处黄素吸收带的丧失。 MAO N和I -PCPA的黄素辅因子之间形成的共价加合物的光谱性质类似于由1-PCPA和哺乳动物的线粒体单胺氧化酶B [Silverman，RB和Zieske，PA（ 1985）生物化学24，2128-2138]。将pH降低至2时，410 nm谱带出现了变色移，表明在灭活后会形成N5-黄素加合物。缺少与哺乳动物形式的MAO A和MAO B中存在的黄素腺嘌呤二核苷酸（FAD）辅因子共价连接的真菌酶MAO N，可以分离并进一步鉴定黄素-灭活剂加合物。将两个13C标签掺入灭活剂[2,3-'3C2] -1-PCPA中，然后使用在线液相色谱/电喷雾电离质谱和核磁共振波谱进行分析，为探索结构提供了一种手段光谱证据支持了1-PCPA灭活剂作为N5-3-氧代-3-苯基丙基-FAD与辅因子的共价结合。

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