Purification and Characterization of Human Uroporphyrinogen 3 Synthase Expressed in Escherichia coli

• Omata Yoshiaki

  Department of Medical Biochemistry, Kurume University School of Medicine

• Sakamoto Hiroshi

  Department of Medical Biochemistry, Kurume University School of Medicine

• Higashimoto Yuichiro

  Department of Medical Biochemistry, Kurume University School of Medicine

• Hayashi Shunsuke

  Department of Medical Biochemistry, Kurume University School of Medicine

• Noguchi Masato

  Department of Medical Biochemistry, Kurume University School of Medicine

書誌事項

タイトル別名

• Purification and Characterization of Human Uroporphyrinogen III Synthase Expressed in <i>Escherichia coli</i>
• Purification and Characterization of Human Uroporphyrinogen III Synthase Expressed in Escherichia coli

抄録

The side-chain asymmetry of physiological porphyrins is produced by the coopera-tive action of hydroxymethylbilane synthase and uroporphyrinogen (uro'gen) III syn-thase. Although the role of uro'gen III synthase is essential for the chemistry of por-phyrin biosynthesis, many aspects, structural as well as mechanical, of uro'gen III synthase have yet to be studied. We report here an expression system in Escherichia coli and a purification procedure for human uro'gen III synthase. The enzyme in the lysate was unstable, but we found that glycerol prevents the activity loss in the lysate. The purified enzyme showed remarkable thermostability, particularly when kept in phosphate buffer containing DTT or EDTA, indicating that the enzyme activity may depend on its oxidation state. Examination of the relationship between the number of Cys residues that are accessible to 5, 5'-dithiobis(2-nitrobenzoic acid) and the remain-ing activity during heat inactivation showed that a particular Cys residue is involved in activity loss. From the crystal structure of human uro'gen III synthase [Mathews et al. (2001) EMBO J. 20, 5832-5839], this Cys residue was considered to be Cys73, which is buried deep inside the enzyme, suggesting that Cys73 of human uro'gen III synthase plays an important role in enzyme activity.

収録刊行物

• The Journal of Biochemistry

  The Journal of Biochemistry 136 (2), 211-220, 2004

  社団法人 日本生化学会

キーワード

• congenital erythropoietic porphyria (CEP)
• porphyrin synthesis
• hydroxymethylbilane synthase
• uroporphyrinogen III synthase

詳細情報

• CRID: 1390282679942767360
• NII論文ID: 130003418166
• NII論文ID: 10016201529
• NII書誌ID: AA00694073
• DOI: 10.1093/jb/mvh111
• COI: 1:CAS:528:DC%2BD2cXpslKrsbw%3D
• ISSN: 17562651
• ISSN: 0021924X
• NDL書誌ID: 7068382
• PubMed: 15496592
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I7068382
• Web Site: http://academic.oup.com/jb/article-pdf/136/2/211/2401952/mvh111.pdf
• Web Site: https://search.jamas.or.jp/link/ui/2006100080
• 本文言語コード: en
• データソース種別:
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  • Crossref
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  • KAKEN
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