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Type: Artigo de periódico
Title: A new member of the aldo-keto reductase family from the plant pathogen Xylella fastidiosa
Author: Rosselli, LK
Oliveira, CLP
Azzoni, AR
Tada, SFS
Catani, CF
Saraiva, AM
Soares, JSM
Medrano, FJ
Torriani, IL
Souza, AP
Abstract: The Xylella fastidiosa genome program generated a large number of gene sequences that belong to pathogenicity, virulence and adaptation categories from this important plant pathogen. One of these genes (XF1729) encodes a protein similar to a superfamily of aldo-keto reductase together with a number of structurally and functionally related NADPH-dependent oxidoreductases. In this work, the similar sequence XF1729 from X. fastidiosa was cloned onto the pET32Xa/LIC vector in order to overexpress a recombinant His-tag fusion protein in Escherichia coli BL21(DE3). The expressed protein in the soluble fraction was purified by immobilized metal affinity chromatography (agarose-IDA-Ni resin). Secondary structure contents were verified by circular dichroism spectroscopy. Small angle Xray scattering (SAXS) measurements furnish general structural parameters and provide a strong indication that the protein has a monomeric form in solution. Also, ab initio calculations show that the protein has some similarities with a previously crystallized aldo-keto reductase protein. The recombinant XF1729 purified to homogeneity catalyzed the reduction of DL-glyceraldehyde (K-cat 2.26 s(-1), K-m 8.20 +/- 0.98 mM) and 2-nitrobenzaldehyde (K-cat 11.74 s(-1), K-m 0.14 +/- 0.04 mM) in the presence of NADPH. The amino acid sequence deduced from XF1729 showed the highest identity (40% or higher) with several functional unknown proteins. Among the identified AKRs, we found approximately 29% of identity with YakC (AKR13), 30 and 28% with AKR11A and AKR11B, respectively. The results establish XF1729 as the new member of AKR family, AKR13B1. Finally, the first characterization by gel filtration chromatography assays indicates that the protein has an elongated shape, which generates an apparent higher molecular weight. The study of this protein is an effort to fight X. fastidiosa, which causes tremendous losses in many economically important plants. (c) 2006 Elsevier Inc. All rights reserved.
Subject: aldo-keto reductase
Xylella fastidiosa
XF1729 protein
XF1729 gene
Country: EUA
Editor: Elsevier Science Inc
Citation: Archives Of Biochemistry And Biophysics. Elsevier Science Inc, v. 453, n. 2, n. 143, n. 150, 2006.
Rights: fechado
Identifier DOI: 10.1016/
Date Issue: 2006
Appears in Collections:Unicamp - Artigos e Outros Documentos

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