The Silent Form of Quinohemoprotein Amine Dehydrogenase from Paracoccus denitrificans

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概要

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• Quinohemoprotein amine dehydrogenase (QH-AmDH) from Paracoccus denitrificans is an αβγ heterotrimeric enzyme catalyzing oxidative deamination of amines with large substitutes, such as butylamine and benzylamine. The smallest γ subunit has cross-linking cysteine tryptophylquinone (CTQ) as a catalytic center. A hemoprotein similar to QH-AmDH in molecular mass, subunit structure, and UV-vis spectral property, but having no enzymatic activity, was isolated. The enzymatically silent form (sQH-AmDH) was activated slowly by the substrates of QH-AmDH, and quickly by reductants, dithionite and dithiothreitol. Electrolysis of sQH-AmDH yielded the active form at potentials more negative than −0.17 V (vs. SHE). The activated protein reacted with a carbonyl reagent, 4-nitrophenylhydrazine, giving a typical spectrum of 4-nitrophenylhydrazone, while sQH-AmDH did not give such a spectrum. The γ subunit of sQH-AmDH showed a sharp peak at 390 nm in UV-vis spectrum clearly distinguishable from that of QH-AmDH. Electrospray ionization mass spectrometric analysis showed that the molecular mass of the γ subunit of sQH-AmDH was larger than that of QH-AmDH by 15.6. The data suggest that the CTQ-like moiety of sQH-AmDH is an oxime. This hypothesis was confirmed by subsequent hydroxylamine treatment of QH-AmDH. QH-AmDH was treated with hydroxylamine yielding an oxime derivative. The UV-vis spectral properties of the γ subunit of hydroxylamine-treated QH-AmDH were identical to those of sQH-AmDH. The hydroxylamine-treated QH-AmDH was also reactivated by butylamine, as in the case of sQH-AmDH.

• 社団法人 日本農芸化学会の論文
• 2009-03-23

著者

• KANO Kenji

  Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University

• IKEDA Tokuji

  Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University

• Kano Kenji

  Kyoto Univ. Kyoto Jpn

• Kano Kenji

  Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University

• Ikeda Tokuji

  Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University

• FUJIEDA Nobutaka

  Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University

• Kano Kenji

  Div. Of Applied Life Sciences Graduate School Of Agriculture Kyoto Univ.

• Mori Megumi

  Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University

• Kano Kenji

  Division Of Applied Life Science Graduate School Of Agriculture Kyoto University

• Mori Megumi

  Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University

• Fujieda Nobutaka

  Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University

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