Molecular cloning of melatonin 2-hydroxylase responsible for 2-hydroxymelatonin production in rice (Oryza sativa)

Journal of Pineal Research
Yeong Byeon, Kyoungwhan Back

Abstract

Although melatonin biosynthetic genes from plants have been cloned, the melatonin catabolism mechanisms remain unclear. To clone the genes responsible for melatonin metabolism, we ectopically expressed 35 full-length cDNAs of rice 2-oxoglutarate-dependent dioxygenase (2-ODD) in Escherichia coli and purified the corresponding recombinant proteins. In vitro 2-ODD assays showed four independent 2-ODD proteins that were able to catalyze melatonin into 2-hydroxymelatonin, exhibiting melatonin 2-hydroxylase (M2H). These M2H proteins had peak activities at pH 8.0 and 30°C. The Km ranged from 121 μm to 371 μm with the Vmax ranging from 1.7 to 18.5 pkat/mg protein, respectively. The M2H enzyme activities were dependent on cofactors such as α-ketoglutarate, ascorbate, and Fe(2+), similar to the 2-ODD enzymes. M2H activity was inhibited by prohexadione-Ca, an inhibitor of 2-ODD, in a dose-dependent manner. M2H activity was high in the roots of rice seedlings, concurrent with high transcription levels of 2-ODD 21, suggesting that 2-ODD 21 was a major gene for M2H activity. Analogous to the high M2H activity in the roots, 2-hydroxymelatonin was found in large quantities in roots treated with melatonin. These results suggest that melatonin w...Continue Reading

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Citations

Oct 27, 2015·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Dun-Xian TanRussel J Reiter
Mar 1, 2016·Frontiers in Plant Science·Rüdiger Hardeland
Mar 2, 2016·Journal of Pineal Research·Hyoung Yool Lee, Kyoungwhan Back
Sep 4, 2015·Journal of Pineal Research·Yeong ByeonKyoungwhan Back
Jun 23, 2015·Journal of Pineal Research·Marino B Arnao, Josefa Hernández-Ruiz
Aug 15, 2015·Journal of Pineal Research·Lucien C ManchesterRussel J Reiter
Nov 2, 2016·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Annia GalanoRussel J Reiter
Nov 22, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Rüdiger Hardeland
May 18, 2018·Journal of Pineal Research·Hyoung Yool Lee, Kyoungwhan Back
Sep 27, 2018·Journal of Pineal Research·Mukesh Kumar KanwarJie Zhou
Jan 12, 2019·Journal of Pineal Research·Sara Muñiz-CalvoJosé M Guillamón
Feb 10, 2019·International Journal of Molecular Sciences·Haoshuang ZhanWeining Song
May 16, 2020·Journal of Experimental Botany·Dun-Xian Tan, Russel J Reiter
Jul 10, 2020·The Plant Journal : for Cell and Molecular Biology·Kyoungwhan Back
Jul 18, 2020·Journal of Integrative Plant Biology·Chengliang SunXianyong Lin
Mar 25, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Yi YangHangun Kim
Aug 1, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Yang YuZhiwei Zhao
Apr 3, 2020·Biomolecules·Ok Jin HwangKyoungwhan Back
May 20, 2021·Journal of Experimental Botany·Stefano NegriFlavia Guzzo
Jul 13, 2021·Frontiers in Plant Science·Susan J Murch, Lauren A E Erland

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