Molecular mode of interaction of plant amine oxidase with the mechanism-based inhibitor 2-butyne-1,4-diamine.
Molecular mode of interaction of plant amine oxidase with the mechanism-based inhibitor 2-butyne-1,4-diamine.
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植物胺氧化酶与基于机制的抑制剂 2-丁炔-1,4-二胺相互作用的分子模式。
DOI：
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
P. Peč
中科院分区：
文献类型：
--
作者：
I. Frébort;M. Šebela;I. Svendsen;S. Hirota;M. Endo;O. Yamauchi;A. Bellelli;K. Lemr;P. Peč
2-丁炔-1，4-二胺(DABI)是一种基于机理的含铜植物胺氧化酶的抑制剂，导致酶失活的次数约为20次。DABI氧化的产物是一种非常活跃的氨基烯，它与酶活性部位的基本亲核基团反应，形成共价结合的吡咯，产生失活的酶。失活的酶在295 nm处有一个新的吸收峰，并产生了与上述试剂处理的吡咯产物光谱相似的对二甲氨基苯甲醛和对二甲氨基肉桂醛的有色衍生物。对二甲氨基苯甲醛吡咯加合物与失活酶的共振拉曼光谱显示出很高的相似性，支持失活产物确实是结合的吡咯的观点。EPR和停流吸收测量表明，在反应的厌氧步骤中已经形成了结合的吡咯，而Topa半醌自由基没有受到影响。含有Dabi结合部位的多肽是通过失活酶的蛋白水解法获得的，经高效液相色谱分离、氨基酸序列分析和MS分析，确定了豌豆胺氧化酶的晶体结构，抑制作用主要是由于高活性的Dabi产物4-氨基-2-丁醛与底物通道入口处的亲核残基结合所致。由于还发现了其他Dabi标记的多肽，并且完全抑制酶后MS没有检测到游离的Dabi产物，因此Dabi产物很可能也与酶表面其他溶剂暴露的亲核残基结合。
2-Butyne-1,4-diamine (DABI) is a mechanism-based inhibitor of copper-containing plant amine oxidases; the number of turnovers that leads to enzyme inactivation is approximately 20. The product of DABI oxidation is a very reactive aminoallene that reacts with an essential nucleophilic group at the enzyme active site, forming a covalently bound pyrrole and producing an inactive enzyme. The inactivated enzyme shows a new absorption maximum at 295 nm and gives coloured derivatives with p-dimethylaminobenzaldehyde and p-dimethylaminocinnamaldehyde that are spectrally similar to the products of pyrrole treated with the above reagents. Resonance Raman spectra of the p-dimethylaminobenzaldehyde adduct of pyrrole and the inactivated enzyme show very high degree of similarity, supporting the idea that the product of inactivation is indeed a bound pyrrole. The bound pyrrole is formed already in the anaerobic step of the reaction, while the topa semiquinone radical is not affected, as shown by the EPR and stopped-flow absorption measurements. Peptides containing the DABI binding site were obtained by proteolysis of inactivated enzyme, isolated by HPLC and analysed by amino acid sequencing and MS. The crystal structure of the amine oxidase from pea has been determined; inhibition is caused mainly by the highly reactive DABI product, 4-amino-2-butynal, binding to a nucleophilic residue at the entrance to the substrate channel. As other DABI labelled peptides were also found and no free DABI product was detected by MS after complete inhibition of the enzyme, it is likely that the DABI product binds also to other solvent exposed nucleophilic residues on the enzyme surface.
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豌豆苗胺氧化酶活性位点铜和托帕醌之间的分子内电子转移速率。
DOI：
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发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Turowski,PN;McGuirl,MA;Dooley,DM
通讯作者：
Dooley,DM
影响因子：
5.7
作者：
Kumar, V;Dooley, DM;Zubak, VM
通讯作者：
Zubak, VM
影响因子：
2.9
作者：
JANES, SM;PALCIC, MM;KLINMAN, JP
通讯作者：
KLINMAN, JP
影响因子：
4.1
作者：
Gellerich, Frank Norbert;Gizatullina, Zemfira;Striggow, Frank
通讯作者：
Striggow, Frank