Studying Protein Complexes by the Yeast Two-hybrid System

Overview

Journal Methods

Specialty Biochemistry

Date 2012 Jul 31

PMID 22841565

Citations 13

Authors

Seesandra V Rajagopala

Patricia Sikorski

J Harry Caufield

Andrey Tovchigrechko

Peter Uetz

Affiliations

Soon will be listed here.

Abstract

Protein complexes are typically analyzed by affinity purification and subsequent mass spectrometric analysis. However, in most cases the structure and topology of the complexes remains elusive from such studies. Here we investigate how the yeast two-hybrid system can be used to analyze direct interactions among proteins in a complex. First we tested all pairwise interactions among the seven proteins of Escherichia coli DNA polymerase III as well as an uncharacterized complex that includes MntR and PerR. Four and seven interactions were identified in these two complexes, respectively. In addition, we review Y2H data for three other complexes of known structure which serve as "gold-standards", namely Varicella Zoster Virus (VZV) ribonucleotide reductase (RNR), the yeast proteasome, and bacteriophage lambda. Finally, we review an Y2H analysis of the human spliceosome which may serve as an example for a dynamic mega-complex.

Citing Articles

Highly Specific Protein Identification by Immunoprecipitation-Mass Spectrometry Using Antifouling Microbeads.

van Andel E, Roosjen M, van der Zanden S, Lange S, Weijers D, Smulders M ACS Appl Mater Interfaces. 2022; .

PMID: 35536557 PMC: 9136845. DOI: 10.1021/acsami.1c22734.

Combined proximity labeling and affinity purification-mass spectrometry workflow for mapping and visualizing protein interaction networks.

Liu X, Salokas K, Weldatsadik R, Gawriyski L, Varjosalo M Nat Protoc. 2020; 15(10):3182-3211.

PMID: 32778839 DOI: 10.1038/s41596-020-0365-x.

Network of Interactions between ZIKA Virus Non-Structural Proteins and Human Host Proteins.

Golubeva V, Nepomuceno T, De Gregoriis G, Mesquita R, Li X, Dash S Cells. 2020; 9(1).

PMID: 31936331 PMC: 7016862. DOI: 10.3390/cells9010153.

Resurrected Protein Interaction Networks Reveal the Innovation Potential of Ancient Whole-Genome Duplication.

Zhang Z, Coenen H, Ruelens P, Hazarika R, Al Hindi T, Oguis G Plant Cell. 2018; 30(11):2741-2760.

PMID: 30333148 PMC: 6305981. DOI: 10.1105/tpc.18.00409.

Mapping the Protein-Protein Interactome Networks Using Yeast Two-Hybrid Screens.

Rajagopala S Adv Exp Med Biol. 2015; 883:187-214.

PMID: 26621469 PMC: 7120425. DOI: 10.1007/978-3-319-23603-2_11.

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