Evaluation of the metal ion requirement of the human deoxyhypusine hydroxylase from HeLa cells using a novel enzyme assay

FEBS Letters
R CsongaH Klier

Abstract

Hypusine synthesis in the eukaryotic initiation factor 5A is a unique two-step posttranslational modification. After deoxyhypusine is generated by the deoxyhypusine synthase, the deoxyhypusine hydroxylase (EC 1.14.99.29) catalyzes the formation of mature hypusine. A rapid assay for monitoring the deoxyhypusine hydroxylase activity was established, employing the oxidative cleavage of the hypusyl residue and subsequent extraction of the generated aldehydes. As metal ion chelators have been reported to inhibit the deoxyhypusine hydroxylase, the mechanism of this inhibition and the effect of transition metal ions on enzyme activity were investigated. A ferric ion appears to be essential for enzymatic activity, the inhibition of which is entirely attributed to the metal ion binding capacity of the chelators.

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Citations

Jul 13, 2002·International Journal of Cancer. Journal International Du Cancer·Paul M J ClementMyung Hee Park
Mar 22, 2006·Parasitology Research·Michael SaeftelAnnette Kaiser
Dec 24, 2005·Proceedings of the National Academy of Sciences of the United States of America·Jong-Hwan ParkMyung Hee Park
Mar 30, 2007·Protein Expression and Purification·Jenq-Kuen HuangLisa Wen
Aug 7, 2009·Journal of Oral Pathology & Medicine : Official Publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology·S-K LeeE-C Kim
Jul 29, 2004·Molecular and Biochemical Parasitology·Ilka M MolitorGabriele M König
Sep 15, 2017·Microbiology and Molecular Biology Reviews : MMBR·Bryan C MounceJohn H Connor
Jul 20, 2006·Cell Biochemistry and Function·Miriam G JasiulionisClaudio M Costa-Neto
Jan 8, 2015·Molecular Pharmacology·Viktoryia SidarovichAlessandro Quattrone

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