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Partial purification and characterization of bovine deoxyhypusine hydroxylase
Author(s) -
Wen Lisa,
Burgess Michelle E,
Huang JenqKuen
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a634-a
Subject(s) - ultrafiltration (renal) , enzyme , biochemistry , affinity chromatography , biology , protein subunit , ion chromatography , gene
The only known protein in eukaryotic cells that contains the unusual amino acid hypusine is the eukaryotic translation initiation factor 5A (eIF5A). Hypusine synthesis in eIF5A consists of two distinct steps involving deoxyhypusine synthase (DHS) and deoxyhypusine hydroxylase (DOHH). Since either DHS or DOHH inhibitors have been shown to exert antiproliferative and antiretroviral effects, they are regarded as potential targets for antiproliferative therapy. Detailed studies directed at DHS or DOHH structures will permit further developing of more specific inhibitors toward attaining better control of hyperproliferative diseases. In this research, we have partially purified bovine DOHH using ultrafiltration and various column chromatography. Our ultrafiltration profiles of DOHH activity suggest that the MW of the enzyme lies between 10 and 50 kDa. The enzyme is most activity at pH 7.4 and the activity is affected by buffer composition. The detection of the DOHH enzyme in multiple fractions eluted under different conditions from ion exchange columns and affinity columns indicates that the DOHH enzyme exists as multiple forms. This is the first report on the existence of multiple forms of DOHH isoenzyme. Supported by National Institute of Health 1R15 GM 60266‐01A1 and the University Research Council, Western Illinois University.