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

FEBS Letters

Auer, Manfred¹  Eckerskorn, Christoph²  Ettmayer, Peter¹  Klier, Hannelore¹  Eder, Jörg¹  Csonga, Robert¹ 

[1] Sandoz Research Institute, Department of Immunodermatology, Brunner Strasse 59, A-1235 Vienna, Austria;Max-Planck-Institute for Biochemistry, Am Klopferspitz 18A, D-82152 Martinsried, Germany

关键词: Eukaryotic initiation factor 5A;    Hypusine;    Posttranslational modification;    Mimosine;    Ciclopirox olamine;    Metal chelation;    CPX;    ciclopirox olamine;    deoxyhypusine synthase;    spermidine dehydrogenase (NAD+;    1-deoxyhypusyl-forming and amino-butyl-transferring);    EDTA;    (ethylenedinitrilo)tetraacetic acid;    DPHP;    4;    6-diphenyl-1-hydroxy-pyridine-2-one;    eIF-5A;    eukaryotic initiation factor 5A;   

DOI  :  10.1016/0014-5793(96)00020-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 the 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 bunding capacity of the chelators.

【 授权许可】

Unknown