Yeast “N”-hybrid systems for protein–protein and drug–protein interaction discovery

Rezwan, Mandana; Auerbach, Daniel

doi:10.1016/j.ymeth.2012.06.006

Cited by 15 publications

(18 citation statements)

References 31 publications

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“…Since its development, the three-hybrid method has been successfully used to identify many protein-small molecule interactions [33, 34]. Very recently, the three-hybrid approach has been used to identify proteins that bind to the drugs erlotinib, atorvastatin, and sulfasalazine [35], the glaucoma therapeutic anecortave acetate [36], and a panel of anti-tuberculosis drugs [37].…”

Section: Methods For Identifying Small Molecule-interacting Cellulamentioning

confidence: 99%

Utilizing Yeast Surface Human Proteome Display Libraries to Identify Small Molecule-Protein Interactions

Bidlingmaier

Liu

2015

Methods in Molecular Biology

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The identification of proteins that interact with small bioactive molecules is a critical but often difficult and time-consuming step in understanding cellular signaling pathways or molecular mechanisms of drug action. Numerous methods for identifying small molecule-interacting proteins have been developed and utilized, including affinity-based purification followed by mass spectrometry analysis, protein microarrays, phage display, and three-hybrid approaches. Although all these methods have been used successfully, there remains a need for additional techniques for analyzing small molecule-protein interactions. A promising method for identifying small molecule-protein interactions is affinity-based selection of yeast surface-displayed human proteome libraries. Large and diverse libraries displaying human protein fragments on the surface of yeast cells have been constructed and subjected to FACS-based enrichment followed by comprehensive exon microarray-based output analysis to identify protein fragments with affinity for small molecule ligands. In a recent example, a proteome-wide search has been successfully carried out to identify cellular proteins binding to the signaling lipids PtdIns(4,5)P2 and PtdIns(3,4,5)P3. Known phosphatidylinositide-binding proteins such as pleckstrin homology domains were identified, as well as many novel interactions. Intriguingly, many novel nuclear phosphatidylinositide-binding proteins were discovered. Although the existence of an independent pool of nuclear phosphatidylinositides has been known about for some time, their functions and mechanism of action remain obscure. Thus, the identification and subsequent study of nuclear phosphatidylinositide-binding proteins is expected to bring new insights to this important biological question. Based on the success with phosphatidylinositides, it is expected that the screening of yeast surface-displayed human proteome libraries will be of general use for the discovery of novel small molecule-protein interactions, thus facilitating the study of cellular signaling pathways and mechanisms of drug action or toxicity.

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Section: Methods For Identifying Small Molecule-interacting Cellulamentioning

confidence: 99%

Utilizing Yeast Surface Human Proteome Display Libraries to Identify Small Molecule-Protein Interactions

Bidlingmaier

Liu

2015

Methods in Molecular Biology

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“…As a consequence, several technologies have been developed, including yeast twohybrid (Y2H), reverse yeast two-hybrid (Rezwan and Auerbach, 2012), bacteria two-hybrid (Stynen et al , 2012), Mammalian Protein-Protein Interaction Trap (MAPPIT) , reverse MAPPIT (Lemmens et al , 2006), bimolecular fluorescence complementation (BiFC), and FRET-and BRETbased methods (Ciruela, 2008;Corbel et al , 2011).…”

Section: Protein-protein Interactions (Ppis)mentioning

confidence: 99%

High‐Throughput Screening of Marine Resources

Hochard

Reininger

Ruchaud

et al. 2014

Outstanding Marine Molecules

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“…Selection at 10 nM, approximately twofold above the LD 50 for parental HCT-116 cells, was conducted to obtain six resistant clones for further analysis. 32 BI 2536, 12, a Polo-like-kinase 1 (PLK1) inhibitor currently undergoing clinical trials, was used to select for resistant HCT-116 colon cancer cells.…”

Section: Transcriptome Sequencing To Identify Mechanisms Of Drug Actimentioning

confidence: 99%

“…These typically are ligands with high affinity 47 to, or substrates that can covalently bind 48 to, a genetically encoded enzyme "hook." 50 The reporter gene amplifies the signal ensuring that unstable targets as well as low affinity transient interactions can be detected. The bifunctional bait upon entry into the nucleus can associate with the hook, creating a preorganized bait scaffold displayed on the promoter region.…”

Section: Yeast Three-hybrid Systemmentioning

confidence: 99%