A Novel Protein Refolding Protocol For The Solubilization And Purification Of Recombinant Peptidoglycan-associated Lipoprotein From Xylella Fastidiosa Overexpressed In Escherichia Coli.

dc.creatorSantos, Clelton A
dc.creatorBeloti, Lilian L
dc.creatorToledo, Marcelo A S
dc.creatorCrucello, Aline
dc.creatorFavaro, Marianna T P
dc.creatorMendes, Juliano S
dc.creatorSantiago, André S
dc.creatorAzzoni, Adriano R
dc.creatorSouza, Anete P
dc.date2012-Apr
dc.date2015-11-27T13:28:11Z
dc.date2015-11-27T13:28:11Z
dc.date.accessioned2018-03-29T01:14:47Z
dc.date.available2018-03-29T01:14:47Z
dc.identifierProtein Expression And Purification. v. 82, n. 2, p. 284-9, 2012-Apr.
dc.identifier1096-0279
dc.identifier10.1016/j.pep.2012.01.010
dc.identifierhttp://www.ncbi.nlm.nih.gov/pubmed/22306742
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/199858
dc.identifier22306742
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1300091
dc.descriptionXylella fastidiosa is a Gram-negative xylem-limited plant pathogenic bacterium responsible for several economically important crop diseases. Here, we present a novel and efficient protein refolding protocol for the solubilization and purification of recombinant X. fastidiosa peptidoglycan-associated lipoprotein (XfPal). Pal is an outer membrane protein that plays important roles in maintaining the integrity of the cell envelope and in bacterial pathogenicity. Because Pal has a highly hydrophobic N-terminal domain, the heterologous expression studies necessary for structural and functional protein characterization are laborious once the recombinant protein is present in inclusion bodies. Our protocol based on the denaturation of the XfPal-enriched inclusion bodies with 8M urea followed by buffer-exchange steps via dialysis proved effective for the solubilization and subsequent purification of XfPal, allowing us to obtain a large amount of relatively pure and folded protein. In addition, XfPal was biochemically and functionally characterized. The method for purification reported herein is valuable for further research on the three-dimensional structure and function of Pal and other outer membrane proteins and can contribute to a better understanding of the role of these proteins in bacterial pathogenicity, especially with regard to the plant pathogen X. fastidiosa.
dc.description82
dc.description284-9
dc.languageeng
dc.relationProtein Expression And Purification
dc.relationProtein Expr. Purif.
dc.rightsfechado
dc.rightsCopyright © 2012 Elsevier Inc. All rights reserved.
dc.sourcePubMed
dc.subjectAmino Acid Sequence
dc.subjectBacterial Proteins
dc.subjectChromatography, Gel
dc.subjectEscherichia Coli
dc.subjectLipoproteins
dc.subjectMolecular Sequence Data
dc.subjectPeptidoglycan
dc.subjectProtein Binding
dc.subjectProtein Refolding
dc.subjectProtein Structure, Quaternary
dc.subjectProtein Structure, Secondary
dc.subjectSequence Homology, Amino Acid
dc.subjectSolubility
dc.subjectXylella
dc.titleA Novel Protein Refolding Protocol For The Solubilization And Purification Of Recombinant Peptidoglycan-associated Lipoprotein From Xylella Fastidiosa Overexpressed In Escherichia Coli.
dc.typeArtículos de revistas


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