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Please use this identifier to cite or link to this item: http://dspace.utalca.cl/handle/1950/8908

Title: Cloning and characterization of Chlorobium tepidum ferritin
Authors: Yevenes, A.E.
Marquez, V.
Watt, R.K..
Keywords: Ferritin
Ferroxidase center
Issue Date: Feb-2011
Citation: BIOCHIMIE Volume: 93 Issue: 2 Pages: 352-360 DOI: 10.1016/j.biochi.2010.10.008
Abstract: The Chlorobium tepidum ferritin (CtFtn) gene was synthesized and cloned into a pET3a expression vector (Novagen). CtFtn was expressed in Escherichia coli and purified to electrophoretic homogeneity. Sequence analysis indicates that all the conserved amino acids required to form the Fe(2+) oxidizing ferroxidase center are present. Ftn is highly conserved from bacteria to humans, each subunit folds into a 4-helical bundle (helices A D), with a long loop connecting helices B and C, plus a fifth short E-helix at the C-terminus. Calculations based on the secondary structure of CtFtn predict that each of these helices forms. However, the sequence of CtFtn shows a much longer C-terminus with a significant number of polar amino acids. Size-exclusion chromatography shows that CtFtn elutes at a size consistent with a 24 -subunit protein cage. Incubation of CtFtn with Fe(2+) produced an increase in the absorbance at 310 nm consistent with the incorporation of iron inside CtFtn. Assays monitoring ferroxidase activity showed that CtFtn possesses ferroxidase activity but it is less active than human H-chain ferritin. Additionally, the iron loading capacity of CtFtn is significantly reduced compared to proteins from other organisms. We propose that the unique extended C-terminus in CtFtn causes the decreased iron loading in CtFtn and possibly influences the slower rate of iron oxidation at the ferroxidase center. (C) 2010 Elsevier Masson SAS. All rights reserved.
Description: Yevenes, AE (Yevenes, Alejandro E.)2,3.....................2. Univ La Frontera, Inst Agroind, Fac Ciencias Agropecuarias & Forestales, Temuco, Chile 3.Univ Talca, Ctr Bioinformat & Simulac Mol, Talca, Chile
URI: http://dspace.utalca.cl/handle/1950/8908
ISSN: 0300-9084
Appears in Collections:Artículos en publicaciones ISI - Universidad de Talca

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