Export of a heterologous cytochrome P450 (CYP105D1) in Escherichia coli is associated with periplasmic accumulation of uroporphyrin

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dc.contributor.author Kelly, Steven L.
dc.contributor.author Akhtar, Kalim M.
dc.contributor.author Kaderbhai, Mustak A.
dc.contributor.author Kaderbhai, Naheed N.
dc.contributor.author Hopper, David J.
dc.date.accessioned 2008-12-16T14:47:04Z
dc.date.available 2008-12-16T14:47:04Z
dc.date.issued 2003-11-14
dc.identifier.citation Kelly , S L , Akhtar , K M , Kaderbhai , M A , Kaderbhai , N N & Hopper , D J 2003 , ' Export of a heterologous cytochrome P450 (CYP105D1) in Escherichia coli is associated with periplasmic accumulation of uroporphyrin ' Journal of Biological Chemistry , pp. 45555-45562 . en
dc.identifier.issn 1083-351X
dc.identifier.other PURE: 93650
dc.identifier.other dspace: 2160/1722
dc.identifier.uri http://hdl.handle.net/2160/1722
dc.identifier.uri http://www.jbc.org/cgi/content/abstract/278/46/45555 en
dc.description M. Kalim Akhtar, Naheed N. Kaderbhai, David J. Hopper, Steven L. Kelly, and Mustak A. Kaderbhai (2003). Export of a heterologous cytochrome P450 (CYP105D1) in Escherichia coli is associated with periplasmic accumulation of uroporphyrin. Journal of Biological Chemistry, 278 (46), 45555-45562. Sponsorship: BBSRC;BBSRC RAE2008 en
dc.description.abstract This report suggests an important physiological role of a CYP in the accumulation of uroporphyrin I arising from catalytic oxidative conversion of uroporphyrinogen I to uroporphyrin I in the periplasm of Escherichia coli cultured in the presence of 5-aminolevulinic acid. A structurally competent Streptomyces griseus CYP105D1 was expressed as an engineered, exportable form in aerobically grown E. coli. Its progressive induction in the presence of 5-aminolevulinic acid-supplemented medium was accompanied by an accumulation of a greater than 100-fold higher amount of uroporphyrin I in the periplasm relative to cells lacking CYP105D1. Expression of a cytoplasm-resident engineered CYP105D1 at a comparative level to the secreted form was far less effective in promoting porphyrin accumulation in the periplasm. Expression at a 10-fold molar excess over the exported CYP105D1 of another periplasmically exported hemoprotein, the globular core of cytochrome b5, did not substitute the role of the periplasmically localized CYP105D1 in promoting porphyrin production. This, therefore, eliminated the possibility that uroporphyrin accumulation is merely a result of increased hemoprotein synthesis. Moreover, in the strain that secreted CYP105D1, uroporphyrin production was considerably reduced by azole-based P450 inhibitors. Production of both holo-CYP105D1 and uroporphyrin was dependent upon 5-aminolevulinic acid, except that at higher concentrations this resulted in a decrease in uroporphyrin. This study suggests that the exported CYP105D1 oxidatively catalyzes periplasmic conversion of uroporphyrinogen I to uroporphyrin I in E. coli. The findings have significant implications in the ontogenesis of human uroporphyria-related diseases. en
dc.format.extent 8 en
dc.language.iso eng
dc.relation.ispartof Journal of Biological Chemistry en
dc.title Export of a heterologous cytochrome P450 (CYP105D1) in Escherichia coli is associated with periplasmic accumulation of uroporphyrin en
dc.type Text en
dc.type.publicationtype Article (Journal) en
dc.identifier.doi http://dx.doi.org/10.1074/jbc.M212685200
dc.contributor.institution Institute of Biological, Environmental and Rural Sciences en
dc.description.status Peer reviewed en


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