Screening of Protein-Protein Interaction Modulators Using BRET-Based Technology

doi:10.1007/978-1-0716-2473-9_12

. 2022:2525:173-183.

doi: 10.1007/978-1-0716-2473-9_12.

Screening of Protein-Protein Interaction Modulators Using BRET-Based Technology

Lucia R Fernández^{1

2}, Jesica Mild^{3

4}, Martin M Edreira^{5

6

7}

Affiliations

¹ Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Química Orgánica, Buenos Aires, Argentina.
² CONICET-Universidad de Buenos Aires, Unidad de Microanálisis y Métodos Físicos Aplicados a la Química Orgánica (UMYMFOR), Buenos Aires, Argentina.
³ Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Química Biológica, Buenos Aires, Argentina.
⁴ CONICET-Universidad de Buenos Aires, Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN), Buenos Aires, Argentina.
⁵ Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Química Biológica, Buenos Aires, Argentina. mme2@pitt.edu.
⁶ CONICET-Universidad de Buenos Aires, Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN), Buenos Aires, Argentina. mme2@pitt.edu.
⁷ Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA. mme2@pitt.edu.

PMID: 35836067
DOI: 10.1007/978-1-0716-2473-9_12

Screening of Protein-Protein Interaction Modulators Using BRET-Based Technology

Lucia R Fernández et al. Methods Mol Biol. 2022.

. 2022:2525:173-183.

doi: 10.1007/978-1-0716-2473-9_12.

Authors

Lucia R Fernández^{1

2}, Jesica Mild^{3

4}, Martin M Edreira^{5

6

7}

Affiliations

¹ Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Química Orgánica, Buenos Aires, Argentina.
² CONICET-Universidad de Buenos Aires, Unidad de Microanálisis y Métodos Físicos Aplicados a la Química Orgánica (UMYMFOR), Buenos Aires, Argentina.
³ Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Química Biológica, Buenos Aires, Argentina.
⁴ CONICET-Universidad de Buenos Aires, Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN), Buenos Aires, Argentina.
⁵ Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Química Biológica, Buenos Aires, Argentina. mme2@pitt.edu.
⁶ CONICET-Universidad de Buenos Aires, Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN), Buenos Aires, Argentina. mme2@pitt.edu.
⁷ Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA. mme2@pitt.edu.

PMID: 35836067
DOI: 10.1007/978-1-0716-2473-9_12

Abstract

Protein-protein interactions (PPIs) play central roles in most molecular mechanisms underlying cellular and biological processes. Within the methods developed to study PPIs is bioluminescence resonance energy transfer (BRET). Taking advantage of this technique, we have set a BRET-based assay that enables the screening of modulators of essential PPIs for Trypanosoma cruzi survival. Considering the complexity of the evaluated mixture, pure chemical compounds or natural extracts, two approaches are described, BRET in living cells or from lysates.

Keywords: Bioluminescence resonance energy transfer (BRET); Drug screening; Inhibitors; Modulators; Protein–protein interaction; Trypanosoma cruzi.

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References

1. Braun P, Gingras AC (2012) History of protein-protein interactions: from egg-white to complex networks. Proteomics 12:1478–1498 - DOI
1. Dimri S, Basu S, De A (2016) Use of BRET to study protein-protein interactions in vitro and in vivo. Methods Mol Biol 1443:57–78. https://doi.org/10.1007/978-1-4939-3724-0_5 - DOI - PubMed
1. Morise H, Shimomura O, Johnson FH, Winant J (1974) Intermolecular energy transfer in the bioluminescent system of aequorea. Biochemistry 13:2656–2662. https://doi.org/10.1021/bi00709a028 - DOI - PubMed
1. Xu Y, Piston DW, Johnson CH (1999) A bioluminescence resonance energy transfer (BRET) system: application to interacting circadian clock proteins. Proc Natl Acad Sci U S A 96:151–156. https://doi.org/10.1073/pnas.96.1.151 - DOI - PubMed - PMC
1. Bacart J, Corbel C, Jockers R et al (2008) The BRET technology and its application to screening assays. Biotechnol J 3:311–324. https://doi.org/10.1002/biot.200700222 - DOI - PubMed

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[1] Braun P, Gingras AC (2012) History of protein-protein interactions: from egg-white to complex networks. Proteomics 12:1478–1498 - DOI

[2] Braun P, Gingras AC (2012) History of protein-protein interactions: from egg-white to complex networks. Proteomics 12:1478–1498 - DOI

[3] Dimri S, Basu S, De A (2016) Use of BRET to study protein-protein interactions in vitro and in vivo. Methods Mol Biol 1443:57–78. https://doi.org/10.1007/978-1-4939-3724-0_5 - DOI - PubMed

[4] Dimri S, Basu S, De A (2016) Use of BRET to study protein-protein interactions in vitro and in vivo. Methods Mol Biol 1443:57–78. https://doi.org/10.1007/978-1-4939-3724-0_5 - DOI - PubMed

[5] Morise H, Shimomura O, Johnson FH, Winant J (1974) Intermolecular energy transfer in the bioluminescent system of aequorea. Biochemistry 13:2656–2662. https://doi.org/10.1021/bi00709a028 - DOI - PubMed

[6] Morise H, Shimomura O, Johnson FH, Winant J (1974) Intermolecular energy transfer in the bioluminescent system of aequorea. Biochemistry 13:2656–2662. https://doi.org/10.1021/bi00709a028 - DOI - PubMed

[7] Xu Y, Piston DW, Johnson CH (1999) A bioluminescence resonance energy transfer (BRET) system: application to interacting circadian clock proteins. Proc Natl Acad Sci U S A 96:151–156. https://doi.org/10.1073/pnas.96.1.151 - DOI - PubMed - PMC

[8] Xu Y, Piston DW, Johnson CH (1999) A bioluminescence resonance energy transfer (BRET) system: application to interacting circadian clock proteins. Proc Natl Acad Sci U S A 96:151–156. https://doi.org/10.1073/pnas.96.1.151 - DOI - PubMed - PMC

[9] Bacart J, Corbel C, Jockers R et al (2008) The BRET technology and its application to screening assays. Biotechnol J 3:311–324. https://doi.org/10.1002/biot.200700222 - DOI - PubMed

[10] Bacart J, Corbel C, Jockers R et al (2008) The BRET technology and its application to screening assays. Biotechnol J 3:311–324. https://doi.org/10.1002/biot.200700222 - DOI - PubMed

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Screening of Protein-Protein Interaction Modulators Using BRET-Based Technology

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Screening of Protein-Protein Interaction Modulators Using BRET-Based Technology

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