Protein-Protein Interactions: Bimolecular Fluorescence Complementation and Cytology Two Hybrid.

Dyuti Purkait, Mohd Ilyas, Krishnamohan Atmakuri

Methods in molecular biology (Clifton, N.J.) 2024

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Identifying protein-protein interactions between machine components of bacterial secretion systems and their cognate substrates is central to delineating how the machines operate to translocate their substrates. Further, establishing which among the machine components and their substrates interact with each other facilitates (i) advancement in our understanding of the architecture and assembly of the machines, (ii) understanding the substrates' translocation routes and mechanisms, and (iii) how the machines and the substrates talk to each other. Currently, though diverse biochemical methods exist in identifying direct and indirect protein-protein interactions, they primarily remain in vitro and can be quite labor intensive. They also may capture/exhibit false-positive interactions because of barrier breakdowns as part of methodology. Thus, adopting novel genetic approaches to help visualize the same in vivo can yield quick, advantageous, reliable, and informative protein-protein interactions data. Here, we describe the easily adoptable bimolecular fluorescence complementation and cytology-based two-hybrid assays to understand the bacterial secretions systems. © 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Dyuti Purkait, Mohd Ilyas, Krishnamohan Atmakuri. Protein-Protein Interactions: Bimolecular Fluorescence Complementation and Cytology Two Hybrid. Methods in molecular biology (Clifton, N.J.). 2024;2715:247-257