Article (Scientific journals)
Towards a universal method for protein refolding: The trimeric beta barrel membrane Omp2a as a test case.
Roussel, Guillaume; Perpete, Eric A.; Matagne, André et al.
2013In Biotechnology and Bioengineering, 110 (2), p. 417-23
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Keywords :
refolding; protein; circular dichroism; 2-methyl-2,4-pentanediol; sodium dodecyl sulfate
Abstract :
[en] It has recently been reported that 2-methyl-2,4-pentanediol (MPD) can modulate the protein-binding properties of sodium dodecyl sulfate (SDS), turning it into a non-denaturing detergent. Indeed both alpha (the lysozyme) and beta (the carbonic anhydrase II) soluble enzymes, as well as a beta membrane protein (PagP) have been successfully refolded into their native form by using this amphiphatic alcohol. In order to support the universal character of our MPD-based technique, we have extended its transferability to the Omp2a trimeric membrane porin. The far-UV circular dichroism signature of Omp2a refolded with our original procedure is identical to that obtained by classical techniques, clearly indicating a proper refolding. Moreover, we show that the optimal SDS/MPD ratio for refolding Omp2a is similar to what has been observed for other types of proteins. While the protocol allows refolding at higher protein concentration (up to 4 mg/mL) and ionic strength (up to 1 M NaCl) than other refolding methods, it is also more efficient at basic pH values and medium temperature (20-40 degrees C). Finally, the key role of the cosolvent was highlighted by a thorough study of the efficiency of MPD analogues, and a high variability was observed, as they can be able or unable to induce refolding at low or high salt concentrations. Biotechnol. Bioeng. 2013; 110: 417-423. (c) 2012 Wiley Periodicals, Inc.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Roussel, Guillaume
Perpete, Eric A.
Matagne, André  ;  Université de Liège - ULiège > Département des sciences de la vie > Enzymologie et repliement des protéines
Tinti, Emmanuel
Michaux, Catherine
Language :
English
Title :
Towards a universal method for protein refolding: The trimeric beta barrel membrane Omp2a as a test case.
Publication date :
2013
Journal title :
Biotechnology and Bioengineering
ISSN :
0006-3592
eISSN :
1097-0290
Publisher :
Wiley - VCH Verlag GmbH & Co., Germany
Volume :
110
Issue :
2
Pages :
417-23
Peer reviewed :
Peer Reviewed verified by ORBi
Commentary :
Copyright (c) 2012 Wiley Periodicals, Inc.
Available on ORBi :
since 28 January 2013

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