Yeast-based assays for detecting protein-protein/drug interactions and their inhibitors

Moosavi, Behrooz; Mousavi, Bibimaryam; Yang, Wen‐Chao; Yang, Guang‐Fu

doi:10.1016/j.ejcb.2017.06.003

Cited by 9 publications

(11 citation statements)

References 149 publications

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“…As a tool for PPI analysis, BioID provides a powerful complement to other established and widely used methods such as the yeast two-hybrid (Y2H) system, co-immunoprecipitation (IP), or pull-down approaches ( Moosavi et al., 2017 ). The BioID method has several significant advantages.…”

Section: Introductionmentioning

confidence: 99%

2C-BioID: An Advanced Two Component BioID System for Precision Mapping of Protein Interactomes

et al. 2018

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SummaryThe modulation of protein-protein interactions (PPIs) is an essential regulatory activity defining diverse cell functions in development and disease. BioID is an unbiased proximity-dependent biotinylation method making use of a biotin-protein ligase fused to a protein of interest and has become an important tool for mapping of PPIs within cellular contexts. We devised an advanced method, 2C-BioID, in which the biotin-protein ligase is kept separate from the protein of interest, until the two are induced to associate by the addition of a dimerizing agent. As proof of principle, we compared the interactomes of lamina-associated polypeptide 2β (LAP2β) with those of lamins A and C, using 2C- and conventional BioID. 2C-BioID greatly enhanced data robustness by facilitating the in silico elimination of non-specific interactors as well as overcoming the problems associated with aberrant protein localization. 2C-BioID therefore significantly strengthens the specificity and reliability of BioID-based interactome analysis, by the more stringent exclusion of false-positives and more efficient intracellular targeting.

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Section: Introductionmentioning

confidence: 99%

2C-BioID: An Advanced Two Component BioID System for Precision Mapping of Protein Interactomes

et al. 2018

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“…Below, we describe briefly selected Y2H variants. More detailed descriptions are provided in several publications [152,153,154]. A schematic diagram of these techniques is shown in Figure 3.…”

Section: State-of-the-art Of the Available Techniques For Detectinmentioning

confidence: 99%

“…It is also possible to first transform a set of different bait constructs in one mating type and a set of preys in the other mating type; then, by generating diploids by mating the two haploid strains, large numbers of binary interactions can be evaluated by growing in selective media (reviewed in [152]).…”

Section: State-of-the-art Of the Available Techniques For Detectinmentioning

confidence: 99%

“…Limitations: Proteins have to enter the nucleus in order to induce the expression of a reporter gene; many false positives and negatives; lack of interaction dynamics and transactivating proteins cannot be properly analyzed. The reasons behind these shortcomings and some minor modifications to solve them have been recently reviewed and some of them will be discussed below [152].…”

Section: State-of-the-art Of the Available Techniques For Detectinmentioning

confidence: 99%

“…This assay is used to identify inhibitors of protein–protein interactions or to select for mutants encoding proteins that have lost their ability to interact, thus providing a high-throughput system to identify inhibitors and key residues mediating protein–protein interactions. Removal of 5-FOA converts this assay to classic Y2H (reviewed in [152]).…”

Section: State-of-the-art Of the Available Techniques For Detectinmentioning

confidence: 99%

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Saccharomyces cerevisiae as a Tool to Investigate Plant Potassium and Sodium Transporters

Locascio

Andrés-Colás

Mulet

et al. 2019

IJMS

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Sodium and potassium are two alkali cations abundant in the biosphere. Potassium is essential for plants and its concentration must be maintained at approximately 150 mM in the plant cell cytoplasm including under circumstances where its concentration is much lower in soil. On the other hand, sodium must be extruded from the plant or accumulated either in the vacuole or in specific plant structures. Maintaining a high intracellular K+/Na+ ratio under adverse environmental conditions or in the presence of salt is essential to maintain cellular homeostasis and to avoid toxicity. The baker’s yeast, Saccharomyces cerevisiae, has been used to identify and characterize participants in potassium and sodium homeostasis in plants for many years. Its utility resides in the fact that the electric gradient across the membrane and the vacuoles is similar to plants. Most plant proteins can be expressed in yeast and are functional in this unicellular model system, which allows for productive structure-function studies for ion transporting proteins. Moreover, yeast can also be used as a high-throughput platform for the identification of genes that confer stress tolerance and for the study of protein–protein interactions. In this review, we summarize advances regarding potassium and sodium transport that have been discovered using the yeast model system, the state-of-the-art of the available techniques and the future directions and opportunities in this field.

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Proteomic applications in identifying protein-protein interactions

Veenstra,

Veenstra

2024

Functional Proteomics

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Yeast-based assays for detecting protein-protein/drug interactions and their inhibitors

Cited by 9 publications

References 149 publications

2C-BioID: An Advanced Two Component BioID System for Precision Mapping of Protein Interactomes

2C-BioID: An Advanced Two Component BioID System for Precision Mapping of Protein Interactomes

Saccharomyces cerevisiae as a Tool to Investigate Plant Potassium and Sodium Transporters

Proteomic applications in identifying protein-protein interactions

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