Literature DB >> 34674177

Using Tandem Affinity Purification to Identify Circadian Clock Protein Complexes from Arabidopsis.

Maria L Sorkin1,2, Dmitri A Nusinow3.   

Abstract

Identification of protein-protein interactions is an effective method of elucidating new roles for circadian clock-associated proteins that can expand beyond the information collected from transcriptional studies and genetic screens. Tandem affinity purification coupled with liquid chromatography mass spectrometry (APMS) utilizes epitope-tagged versions of your protein of interest to co-precipitate direct and indirect protein partners. Here, we provide a protocol and suggestions for proper design of 6x-His-3x-FLAG-tagged clock proteins and isolation of protein-protein interactions using two immunoprecipitation steps for increased specificity.
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Entities:  

Keywords:  Affinity purification; Circadian clock; Mass spectrometry; Protein-protein interactions

Mesh：

Substances：

Year:  2022        PMID： 34674177     DOI： 10.1007/978-1-0716-1912-4_15

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


  4 in total

1.  An improved toolbox to unravel the plant cellular machinery by tandem affinity purification of Arabidopsis protein complexes.

Authors:  Jelle Van Leene; Dominique Eeckhout; Bernard Cannoot; Nancy De Winne; Geert Persiau; Eveline Van De Slijke; Leen Vercruysse; Maarten Dedecker; Aurine Verkest; Klaas Vandepoele; Lennart Martens; Erwin Witters; Kris Gevaert; Geert De Jaeger
Journal:  Nat Protoc       Date:  2014-12-18       Impact factor: 13.491

Review 2.  Circadian Rhythms in Plants.

Authors:  Nicky Creux; Stacey Harmer
Journal:  Cold Spring Harb Perspect Biol       Date:  2019-09-03       Impact factor: 10.005

3.  GSyellow, a Multifaceted Tag for Functional Protein Analysis in Monocot and Dicot Plants.

Authors:  Nienke Besbrugge; Jelle Van Leene; Dominique Eeckhout; Bernard Cannoot; Shubhada R Kulkarni; Nancy De Winne; Geert Persiau; Eveline Van De Slijke; Michiel Bontinck; Stijn Aesaert; Francis Impens; Kris Gevaert; Daniel Van Damme; Mieke Van Lijsebettens; Dirk Inzé; Klaas Vandepoele; Hilde Nelissen; Geert De Jaeger
Journal:  Plant Physiol       Date:  2018-04-20       Impact factor: 8.340

4.  Identification of Evening Complex Associated Proteins in Arabidopsis by Affinity Purification and Mass Spectrometry.

Authors:  He Huang; Sophie Alvarez; Rebecca Bindbeutel; Zhouxin Shen; Michael J Naldrett; Bradley S Evans; Steven P Briggs; Leslie M Hicks; Steve A Kay; Dmitri A Nusinow
Journal:  Mol Cell Proteomics       Date:  2015-11-06       Impact factor: 5.911
  4 in total

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