Literature DB >> 24943314

Simplified protein purification using an autoprocessing, inducible enzyme tag.

Aimee Shen1.   

Abstract

The development of affinity tags has greatly simplified protein purification procedures. A variety of affinity tags are now available to improve expression, solubility, and/or tag removal. In this chapter, we describe a method for purifying recombinant proteins expressed in Escherichia coli that uses a highly specific, inducible, C-terminal autoprocessing protease tag. This method streamlines affinity purification, cleavage, and tag separation into a one-step purification procedure, avoiding the need to remove fusion tags from target proteins with exogenous proteases. In addition to accelerating protein purification, we show that this method can enhance the expression, stability, and solubility of select proteins.

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Year:  2014        PMID: 24943314      PMCID: PMC4377278          DOI: 10.1007/978-1-4939-1034-2_5

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


  11 in total

Review 1.  Making the most of affinity tags.

Authors:  David S Waugh
Journal:  Trends Biotechnol       Date:  2005-06       Impact factor: 19.536

Review 2.  Current strategies for the use of affinity tags and tag removal for the purification of recombinant proteins.

Authors:  José Arnau; Conni Lauritzen; Gitte E Petersen; John Pedersen
Journal:  Protein Expr Purif       Date:  2005-12-28       Impact factor: 1.650

3.  Structural and molecular mechanism for autoprocessing of MARTX toxin of Vibrio cholerae at multiple sites.

Authors:  Katerina Prochazkova; Ludmilla A Shuvalova; George Minasov; Zdenek Voburka; Wayne F Anderson; Karla J F Satchell
Journal:  J Biol Chem       Date:  2009-07-20       Impact factor: 5.157

4.  BirA enzyme: production and application in the study of membrane receptor-ligand interactions by site-specific biotinylation.

Authors:  C A O'callaghan; M F Byford; J R Wyer; B E Willcox; B K Jakobsen; A J McMichael; J I Bell
Journal:  Anal Biochem       Date:  1999-01-01       Impact factor: 3.365

5.  Recombinant production of cathelicidin-derived antimicrobial peptides in Escherichia coli using an inducible autocleaving enzyme tag.

Authors:  Oliver Wright; Tatsuya Yoshimi; Alan Tunnacliffe
Journal:  N Biotechnol       Date:  2011-11-09       Impact factor: 5.079

6.  Large-scale expression, refolding, and purification of the catalytic domain of human macrophage metalloelastase (MMP-12) in Escherichia coli.

Authors:  A A Parkar; M D Stow; K Smith; A K Panicker; J P Guilloteau; R Jupp; S J Crowe
Journal:  Protein Expr Purif       Date:  2000-11       Impact factor: 1.650

Review 7.  myo-Inositol-1,2,3,4,5,6-hexakisphosphate.

Authors:  Victor Raboy
Journal:  Phytochemistry       Date:  2003-11       Impact factor: 4.072

Review 8.  Inositol derivatives: evolution and functions.

Authors:  Robert H Michell
Journal:  Nat Rev Mol Cell Biol       Date:  2008-02       Impact factor: 94.444

9.  Simplified, enhanced protein purification using an inducible, autoprocessing enzyme tag.

Authors:  Aimee Shen; Patrick J Lupardus; Montse Morell; Elizabeth L Ponder; A Masoud Sadaghiani; K Christopher Garcia; Matthew Bogyo
Journal:  PLoS One       Date:  2009-12-02       Impact factor: 3.240

10.  Mechanistic and structural insights into the proteolytic activation of Vibrio cholerae MARTX toxin.

Authors:  Aimee Shen; Patrick J Lupardus; Victoria E Albrow; Andrew Guzzetta; James C Powers; K Christopher Garcia; Matthew Bogyo
Journal:  Nat Chem Biol       Date:  2009-07       Impact factor: 15.040

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