Abstract:
:A gene (yacK) encoding a putative multicopper oxidase (MCO) was cloned from Escherichia coli, and the expressed enzyme was demonstrated to exhibit phenoloxidase and ferroxidase activities. The purified protein contained six copper atoms per polypeptide chain and displayed optical and electron paramagnetic resonance (EPR) spectra consistent with the presence of type 1, type 2, and type 3 copper centers. The strong optical A(610) (E(610) = 10,890 M(-1) cm(-1)) and copper stoichiometry were taken as evidence that, similar to ceruloplasmin, the enzyme likely contains multiple type 1 copper centers. The addition of copper led to immediate and reversible changes in the optical and EPR spectra of the protein, as well as decreased thermal stability of the enzyme. Copper addition also stimulated both the phenoloxidase and ferroxidase activities of the enzyme, but the other metals tested had no effect. In the presence of added copper, the enzyme displayed significant activity against two of the phenolate siderophores utilized by E. coli for iron uptake, 2,3-dihydroxybenzoate and enterobactin, as well as 3-hydroxyanthranilate, an iron siderophore utilized by Saccharomyces cerevisiae. Oxidation of enterobactin produced a colored precipitate suggestive of the polymerization reactions that characterize microbial melanization processes. As oxidation should render the phenolate siderophores incapable of binding iron, yacK MCO activity could influence levels of free iron in the periplasm in response to copper concentration. This mechanism may explain, in part, how yacK MCO moderates the sensitivity of E. coli to copper.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Kim C,Lorenz WW,Hoopes JT,Dean JFdoi
10.1128/JB.183.16.4866-4875.2001keywords:
subject
Has Abstractpub_date
2001-08-01 00:00:00pages
4866-75issue
16eissn
0021-9193issn
1098-5530journal_volume
183pub_type
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journal_title:Journal of bacteriology
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doi:10.1128/JB.117.3.1017-1022.1974
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更新日期:1982-07-01 00:00:00
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更新日期:2005-12-01 00:00:00
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更新日期:1997-06-01 00:00:00
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pub_type: 杂志文章
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更新日期:2007-02-01 00:00:00
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更新日期:1970-05-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.113.3.1363-1372.1973
更新日期:1973-03-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.178.7.1872-1880.1996
更新日期:1996-04-01 00:00:00
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更新日期:1990-08-01 00:00:00
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更新日期:1972-09-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1965-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1989-09-01 00:00:00
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pub_type: 杂志文章
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更新日期:1995-08-01 00:00:00
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pub_type: 杂志文章
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更新日期:2010-06-01 00:00:00
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pub_type: 杂志文章
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更新日期:1965-05-01 00:00:00
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pub_type: 杂志文章
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更新日期:1973-07-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1989-06-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.153.1.84-92.1983
更新日期:1983-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:2005-05-01 00:00:00
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pub_type: 杂志文章
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更新日期:1972-10-01 00:00:00