Article (Scientific journals)
Purification, refolding and characterization of the trimeric Omp2a outer membrane porin from Brucella melitensis.
Roussel, G; Matagne, André; De Bolle, X et al.
2012In Protein Expression and Purification, 83 (2), p. 198-204
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Keywords :
circular dichroism; membrane protein; porin; refolding; fluorescence
Abstract :
[en] Brucella melitensis is a gram-negative bacteria known to cause brucellosis and to produce severe infections in humans. Whilst brucella's outer membrane proteins have been extensively studied due to their potential role as antigens or virulence factors, their function is still poorly understood at the structural level, as the 3D structure of Brucella beta-barrel membrane proteins are still unknown. In this context, the B. melitensis trimeric Omp2a porin has been overexpressed and refolded in n-dodecyl-beta-d-maltopyranoside. We here show that this refolding process is insensitive to urea but is temperature- and ionic strength-dependent. Reassembled species were characterized by fluorescence, size-exclusion chromatography and circular dichroism. A refolding mechanism is proposed, suggesting that Omp2a first refolds under a monomeric form and then self-associates into a trimeric state. This first complete in vitro refolding of a membrane protein from B. melitensis shall eventually lead to functional and 3D structure determination.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Roussel, G
Matagne, André  ;  Université de Liège - ULiège > Département des sciences de la vie > Enzymologie et repliement des protéines
De Bolle, X
Perpete, EA
Michaux, Catherine
Language :
English
Title :
Purification, refolding and characterization of the trimeric Omp2a outer membrane porin from Brucella melitensis.
Publication date :
2012
Journal title :
Protein Expression and Purification
ISSN :
1046-5928
eISSN :
1096-0279
Publisher :
Academic Press, Orlando, United States - Florida
Volume :
83
Issue :
2
Pages :
198-204
Peer reviewed :
Peer Reviewed verified by ORBi
Commentary :
Copyright (c) 2012 Elsevier Inc. All rights reserved.
Available on ORBi :
since 11 May 2012

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