An automatable screen for the rapid identification of proteins amenable to refolding

Cowieson, Nathan P., Wensley, Beth, Listwan, Pawel, Hume, David A., Kobe, Bostjan and Martin, Jennifer L. (2006) An automatable screen for the rapid identification of proteins amenable to refolding. Proteomics, 6 6: 1750-1757.


Author Cowieson, Nathan P.
Wensley, Beth
Listwan, Pawel
Hume, David A.
Kobe, Bostjan
Martin, Jennifer L.
Title An automatable screen for the rapid identification of proteins amenable to refolding
Journal name Proteomics   Check publisher's open access policy
ISSN 1615-9853
Publication date 2006
Sub-type Article (original research)
DOI 10.1002/pmic.200500056
Volume 6
Issue 6
Start page 1750
End page 1757
Total pages 8
Editor Michael Dunn
Place of publication Germany
Publisher Wiley-V C H Verlag GmbH & co KGaA
Collection year 2006
Language eng
Subject C1
270199 Biochemistry and Cell Biology not elsewhere classified
780105 Biological sciences
Abstract Insoluble expression of heterologous proteins in Escherichia coli is a major bottleneck of many structural genomics and high-throughput protein biochemistry projects. Many of these proteins may be amenable to refolding, but their identification is hampered by a lack of high-throughput methods. We have developed a matrix-assisted refolding approach in which correctly folded proteins are distinguished from misfolded proteins by their elution from affinity resin under nondenaturing conditions. Misfolded proteins remain adhered to the resin, presumably via hydrophobic interactions. The assay can be applied to insoluble proteins on an individual basis but is particularly well suited for high-throughput applications because it is rapid, automatable and has no rigorous sample preparation requirements. The efficacy of the screen is demonstrated on small-scale expression samples for 15 proteins. Refolding is then validated by large-scale expressions using SEC and circular dichroism.
Keyword Biochemical Research Methods
protein expression screening
High-throughput
Protein Expression Screening
Protein Refolding
Structural Genomics
Structural Genomics Projects
Secondary Structure
Folding Screen
Expression
Purification
Proteomics
Q-Index Code C1

 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 10 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 11 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Access Statistics: 76 Abstract Views  -  Detailed Statistics
Created: Wed, 15 Aug 2007, 08:24:34 EST