Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/195800
Type: Artigo de periódico
Title: Expression And Purification Of A Small Heat Shock Protein From The Plant Pathogen Xylella Fastidiosa.
Author: Azzoni, Adriano R
Tada, Susely F S
Rosselli, Luciana K
Paula, Débora P
Catani, Cleide F
Sabino, Adão A
Barbosa, João A R G
Guimarães, Beatriz G
Eberlin, Marcos N
Medrano, Francisco J
Souza, Anete P
Abstract: The small heat shock proteins (smHSPs) belong to a family of proteins that function as molecular chaperones by preventing protein aggregation and are also known to contain a conserved region termed alpha-crystallin domain. Here, we report the expression, purification, and partial characterization of a novel smHSP (HSP17.9) from the phytopathogen Xylella fastidiosa, causal agent of the citrus variegated chlorosis (CVC). The gene was cloned into a pET32-Xa/LIC vector to over-express the protein coupled with fusion tags in Escherichia coli BL21(DE3). The expressed HSP17.9 was purified by immobilized metal affinity chromatography (IMAC) and had its identity determined by mass spectrometry (MALDI-TOF). The correct folding of the purified recombinant protein was verified by circular dichroism spectroscopy. Finally, the HSP17.9 protein also proved to efficiently prevent induced aggregation of insulin, strongly indicating a chaperone-like activity.
Subject: Amino Acid Sequence
Circular Dichroism
Escherichia Coli
Gene Expression
Genetic Vectors
Heat-shock Proteins
Insulin
Molecular Sequence Data
Oligonucleotides
Plasmids
Recombinant Proteins
Spectrometry, Mass, Matrix-assisted Laser Desorption-ionization
Structure-activity Relationship
Time Factors
Transformation, Bacterial
Xylella
Rights: fechado
Identifier DOI: 
Address: http://www.ncbi.nlm.nih.gov/pubmed/14711518
Date Issue: 2004
Appears in Collections:Artigos e Materiais de Revistas Científicas - Unicamp

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