Literature DB >> 21215316

The epitope for the polyol-responsive monoclonal antibody 8RB13 is in the flap-domain of the beta-subunit of bacterial RNA polymerase and can be used as an epitope tag for immunoaffinity chromatography.

Elizabeth S Stalder1, Lauren H Nagy, Pilar Batalla, Terrance M Arthur, Nancy E Thompson, Richard R Burgess.   

Abstract

Polyol-responsive monoclonal antibodies (PR-mAbs) are useful for the purification of proteins in an easy, one step immunoaffinity step. These antibodies allow for gentle purification of proteins and protein complexes using a combination of a low molecular weight polyhydroxylated compound (polyol) and a nonchaotrophic salt in the eluting buffer. mAb 8RB13 has been characterized as one of these PR-mAbs and has been used to purify RNA polymerase from five species of bacteria. Here the epitope for 8RB13 has been identified as PEEKLLRAIFGEKAS, a sequence that is highly conserved in the β-subunit of bacterial RNA polymerase. This sequence is located in the "beta-flap" domain of RNA polymerase (and essentially comprises the "flap-tip helix"), an important binding site for sigma70. This location explains why only the core RNAP is purified using this mAb. This amino acid sequence has been developed into an epitope tag that can be used to purify a target protein from either bacterial or eukaryotic cells when genetically fused to a protein of interest.
Copyright © 2011 Elsevier Inc. All rights reserved.

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Year:  2011        PMID: 21215316      PMCID: PMC3248238          DOI: 10.1016/j.pep.2010.12.011

Source DB:  PubMed          Journal:  Protein Expr Purif        ISSN: 1046-5928            Impact factor:   1.650


  14 in total

1.  Mapping protein-protein interaction domains using ordered fragment ladder far-western analysis of hexahistidine-tagged fusion proteins.

Authors:  R R Burgess; T M Arthur; B C Pietz
Journal:  Methods Enzymol       Date:  2000       Impact factor: 1.600

2.  GroEL-GroES solubilizes abundantly expressed xylulokinase in Escherichia coli.

Authors:  S Bu; P W K Tsang; R Z Fu
Journal:  J Appl Microbiol       Date:  2005       Impact factor: 3.772

3.  Isolation and characterization of a polyol-responsive monoclonal antibody useful for gentle purification of Escherichia coli RNA polymerase.

Authors:  N E Thompson; D A Hager; R R Burgess
Journal:  Biochemistry       Date:  1992-08-04       Impact factor: 3.162

4.  Identification of polyol-responsive monoclonal antibodies for use in immunoaffinity chromatography.

Authors:  N E Thompson; R R Burgess
Journal:  Curr Protoc Mol Biol       Date:  2001-05

Review 5.  Identification, production, and use of polyol-responsive monoclonal antibodies for immunoaffinity chromatography.

Authors:  Nancy E Thompson; Katherine M Foley; Elizabeth S Stalder; Richard R Burgess
Journal:  Methods Enzymol       Date:  2009       Impact factor: 1.600

6.  Localization of a sigma70 binding site on the N terminus of the Escherichia coli RNA polymerase beta' subunit.

Authors:  T M Arthur; R R Burgess
Journal:  J Biol Chem       Date:  1998-11-20       Impact factor: 5.157

7.  Epitope mapping using histidine-tagged protein fragments: application to Escherichia coli RNA polymerase sigma 70.

Authors:  L Rao; D P Jones; L H Nguyen; S A McMahan; R R Burgess
Journal:  Anal Biochem       Date:  1996-10-15       Impact factor: 3.365

8.  A hydrophobic patch on the flap-tip helix of E.coli RNA polymerase mediates sigma(70) region 4 function.

Authors:  Kati Geszvain; Tanja M Gruber; Rachel A Mooney; Carol A Gross; Robert Landick
Journal:  J Mol Biol       Date:  2004-10-22       Impact factor: 5.469

9.  Molecular evolution of multisubunit RNA polymerases: structural analysis.

Authors:  William J Lane; Seth A Darst
Journal:  J Mol Biol       Date:  2009-11-03       Impact factor: 5.469

10.  Immunoaffinity purification and characterization of RNA polymerase from Shewanella oneidensis.

Authors:  Mitchell D Probasco; Nancy E Thompson; Richard R Burgess
Journal:  Protein Expr Purif       Date:  2007-04-10       Impact factor: 1.650

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  6 in total

1.  Co-ordination between BrlA regulation and secretion of the oxidoreductase FmqD directs selective accumulation of fumiquinazoline C to conidial tissues in Aspergillus fumigatus.

Authors:  Fang Yun Lim; Brian Ames; Christopher T Walsh; Nancy P Keller
Journal:  Cell Microbiol       Date:  2014-04-15       Impact factor: 3.715

2.  Automated operation of immiscible filtration assisted by surface tension (IFAST) arrays for streamlined analyte isolation.

Authors:  Scott M Berry; Keil J Regehr; Benjamin P Casavant; David J Beebe
Journal:  J Lab Autom       Date:  2012-09-26

3.  Weak protein-protein interactions revealed by immiscible filtration assisted by surface tension.

Authors:  Scott M Berry; Emily N Chin; Shawn S Jackson; Lindsay N Strotman; Mohit Goel; Nancy E Thompson; Caroline M Alexander; Shigeki Miyamoto; Richard R Burgess; David J Beebe
Journal:  Anal Biochem       Date:  2013-11-09       Impact factor: 3.365

4.  Evaluation of the antibacterial activity of patchouli oil.

Authors:  Xian Yang; Xue Zhang; Shui-Ping Yang; Wei-Qi Liu
Journal:  Iran J Pharm Res       Date:  2013       Impact factor: 1.696

5.  AirJump: Using Interfaces to Instantly Perform Simultaneous Extractions.

Authors:  Scott M Berry; Hannah M Pezzi; Alex J LaVanway; David J Guckenberger; Meghan A Anderson; David J Beebe
Journal:  ACS Appl Mater Interfaces       Date:  2016-06-10       Impact factor: 9.229

6.  Ms1, a novel sRNA interacting with the RNA polymerase core in mycobacteria.

Authors:  Jarmila Hnilicová; Jitka Jirát Matějčková; Michaela Šiková; Jiří Pospíšil; Petr Halada; Josef Pánek; Libor Krásný
Journal:  Nucleic Acids Res       Date:  2014-09-12       Impact factor: 16.971

  6 in total

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