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PMID: 12824478 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S. Review

Crystal structures of fusion proteins with large-affinity tags.

Protein science : a publication of the Protein Society ·Vol. 12 ·No. 7 ·2003-07-00 ·Pages 1313-22

Smyth DR, Mrozkiewicz MK, McGrath WJ, Listwan P, Kobe B

Abstract

The fusion of a protein of interest to a large-affinity tag, such as the maltose-binding protein (MBP), thioredoxin (TRX), or glutathione-S-transferase (GST), can be advantageous in terms of increased expression, enhanced solubility, protection from proteolysis, improved folding, and protein purification via affinity chromatography. Unfortunately, crystal growth is hindered by the conformational heterogeneity induced by the fusion tag, requiring that the tag is removed by a potentially problematic cleavage step. The first three crystal structures of fusion proteins with large-affinity tags have been reported recently. All three structures used a novel strategy to rigidly fuse the protein of interest to MBP via a short three- to five-amino acid spacer. This strategy has the potential to aid structure determination of proteins that present particular experimental challenges and are not conducive to more conventional crystallization strategies (e.g., membrane proteins). Structural genomics initiatives may also benefit from this approach as a way to crystallize problematic proteins of significant interest.

MeSH Terms
Affinity Labels/chemistry Carrier Proteins/chemistry Crystallography, X-Ray Dimerization Genomics/methods Glutathione Transferase/chemistry Maltose-Binding Proteins Membrane Proteins/chemistry Protein Conformation Recombinant Fusion Proteins/chemistry,isolation & purification Thioredoxins/chemistry
Chemicals
Affinity Labels Carrier Proteins Maltose-Binding Proteins Membrane Proteins Recombinant Fusion Proteins Thioredoxins Glutathione Transferase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Smyth Douglas R
Department of Biochemistry and Molecular Biology, Institute for Molecular Bioscience, and Special Research Centre for Functional and Applied Genomics, University of Queensland, St Lucia, Queensland 4072, Australia.
Mrozkiewicz Marek K
McGrath William J
Listwan Pawel
Kobe Bostjan
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Article Info
Journal
Protein science : a publication of the Protein Society
Abbr.
Protein Sci
ISSN
0961-8368
DOI
10.1110/ps.0243403
Published
2003-07-00
Pages
1313-22
Language
English
Region
United States
NLM ID
9211750
PMCID
PMC2323919
Subset
IM
External Links
PubMed source DOI: 10.1110/ps.0243403 PMC Full Text View journal details
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