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Cytotrap: An Innovative Approach for Protein–Protein Interaction Studies for Cytoplasmic Proteins

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Protein-Protein Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2690))

Abstract

Protein–protein interaction mapping has gained immense importance in understanding protein functions in diverse biological pathways. There are various in vivo and in vitro techniques associated with the protein–protein interaction studies but generally, the focus is confined to understanding the protein interaction in the nucleus of the cell, and thus it limits the availability to explore protein interactions that are happening in the cytoplasm of the cell. Since posttranslational modification is a crucial step in signaling pathways and cellular protein interactions harnessing the cytoplasmic protein and evaluating the interaction in the cytoplasm, this protocol will provide more information about studying these types of protein interactions. Cytotrap is a type of yeast-two-hybrid system that differs in its ability to anchor along the membrane, thus directing the protein of interest to anchor along the membrane through the myristoylation signaling unit. The vector containing the target protein contains the myristoylation unit, called the prey, and the bait unit contains the protein of interest as a fusion with the hSos protein. In an event of interaction between the target and the protein of interest, the hSos protein unit will be localized to the membrane and the GDP/GTP exchange unit will trigger the activation of the Ras pathway that leads to the survival of the temperature-sensitive yeast strain at a higher temperature.

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Acknowledgments

This research was funded by the National Science Foundation (IOS-2038872).

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Authors and Affiliations

  1. Department of Biology, University of Alabama, Birmingham, AL, USA

    Binoop Mohan, Doni Thingujam & Karolina M. Pajerowska-Mukhtar

Authors
  1. Binoop Mohan
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  2. Doni Thingujam
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  3. Karolina M. Pajerowska-Mukhtar
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Corresponding author

Correspondence to Karolina M. Pajerowska-Mukhtar .

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Editors and Affiliations

  1. Department of Biology, University of Alabama at Birmingham, Birmingham, AL, USA

    Shahid Mukhtar

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Mohan, B., Thingujam, D., Pajerowska-Mukhtar, K.M. (2023). Cytotrap: An Innovative Approach for Protein–Protein Interaction Studies for Cytoplasmic Proteins. In: Mukhtar, S. (eds) Protein-Protein Interactions. Methods in Molecular Biology, vol 2690. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3327-4_2

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  • DOI: https://doi.org/10.1007/978-1-0716-3327-4_2

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3326-7

  • Online ISBN: 978-1-0716-3327-4

  • eBook Packages: Springer Protocols

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