Measuring NLR Oligomerization III: Detection of NLRP3 and NLRC4 Complex by Bioluminescence Resonance Energy Transfer

doi:10.1007/978-1-0716-3350-2_6

. 2023:2696:93-103.

doi: 10.1007/978-1-0716-3350-2_6.

Measuring NLR Oligomerization III: Detection of NLRP3 and NLRC4 Complex by Bioluminescence Resonance Energy Transfer

Fátima Martín-Sánchez¹, Alejandro Peñín-Franch², Diego Angosto-Bazarra², Ana Tapia-Abellán², Vincent Compan^{3

4}, Pablo Pelegrín^{5

6}

Affiliations

¹ Inflammation and Experimental Surgery Unit, Institute for Bio-Health Research of Murcia, Clinical University Hospital Virgen de la Arrixaca, Murcia, Spain.
² Biomedical Research Institute of Murcia, Murcia, Spain.
³ Institut de Génomique Fonctionnelle, Labex ICST, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Université Montpellier, Montpellier, France.
⁴ Institut National de la Santé et de la Recherche Médicale Unité 1191, Montpellier, France.
⁵ Biomedical Research Institute of Murcia, Murcia, Spain. pablopel@um.es.
⁶ Department of Bioquimestry and Molecular Biology B and Immunology, Faculty of Medicine, University of Murcia, Murcia, Spain. pablopel@um.es.

PMID: 37578717
DOI: 10.1007/978-1-0716-3350-2_6

Measuring NLR Oligomerization III: Detection of NLRP3 and NLRC4 Complex by Bioluminescence Resonance Energy Transfer

Fátima Martín-Sánchez et al. Methods Mol Biol. 2023.

. 2023:2696:93-103.

doi: 10.1007/978-1-0716-3350-2_6.

Authors

Fátima Martín-Sánchez¹, Alejandro Peñín-Franch², Diego Angosto-Bazarra², Ana Tapia-Abellán², Vincent Compan^{3

4}, Pablo Pelegrín^{5

6}

Affiliations

¹ Inflammation and Experimental Surgery Unit, Institute for Bio-Health Research of Murcia, Clinical University Hospital Virgen de la Arrixaca, Murcia, Spain.
² Biomedical Research Institute of Murcia, Murcia, Spain.
³ Institut de Génomique Fonctionnelle, Labex ICST, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Université Montpellier, Montpellier, France.
⁴ Institut National de la Santé et de la Recherche Médicale Unité 1191, Montpellier, France.
⁵ Biomedical Research Institute of Murcia, Murcia, Spain. pablopel@um.es.
⁶ Department of Bioquimestry and Molecular Biology B and Immunology, Faculty of Medicine, University of Murcia, Murcia, Spain. pablopel@um.es.

PMID: 37578717
DOI: 10.1007/978-1-0716-3350-2_6

Abstract

Bioluminescent resonance energy transfer (BRET) is a natural phenomenon resulting from a non-radiative energy transfer between a bioluminescent donor (Renilla luciferase) and a fluorescent protein acceptor. BRET signal is dependent on the distance and the orientation between the donor and the acceptor and could be used to study protein-protein interactions and conformational changes within proteins at real-time in living cells. This protocol describes the use of BRET technique to study NLRP3 oligomerization in living cells before and during NLRP3 inflammasome activation.

Keywords: BRET; Bioluminescence; NLRC4; NLRP3; Sensor.

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References

1. Pfleger KDG, Eidne KA (2006) Illuminating insights into protein-protein interactions using bioluminescence resonance energy transfer (BRET). Nat Methods 3:165–174 - DOI - PubMed
1. Pfleger KDG, Seeber RM, Eidne KA (2006) Bioluminescence resonance energy transfer (BRET) for the real-time detection of protein-protein interactions. Nat Protoc 1:337–345 - DOI - PubMed
1. Xu Y, Kanauchi A, von Arnim AG, Piston DW, Johnson CH (2003) Bioluminescence resonance energy transfer: monitoring protein-protein interactions in living cells. Methods Enzymol 360:289–301 - DOI - PubMed
1. Wu PG, Brand L (1994) Resonance energy transfer: methods and applications. Anal Biochem 218:1–13 - DOI - PubMed
1. Siddiqui S, Cong W-N, Daimon CM, Martin B, Maudsley S (2013) BRET biosensor analysis of receptor tyrosine kinase functionality. Front Endocrinol (Lausanne) 4:46 - DOI - PubMed

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[1] Pfleger KDG, Eidne KA (2006) Illuminating insights into protein-protein interactions using bioluminescence resonance energy transfer (BRET). Nat Methods 3:165–174 - DOI - PubMed

[2] Pfleger KDG, Eidne KA (2006) Illuminating insights into protein-protein interactions using bioluminescence resonance energy transfer (BRET). Nat Methods 3:165–174 - DOI - PubMed

[3] Pfleger KDG, Seeber RM, Eidne KA (2006) Bioluminescence resonance energy transfer (BRET) for the real-time detection of protein-protein interactions. Nat Protoc 1:337–345 - DOI - PubMed

[4] Pfleger KDG, Seeber RM, Eidne KA (2006) Bioluminescence resonance energy transfer (BRET) for the real-time detection of protein-protein interactions. Nat Protoc 1:337–345 - DOI - PubMed

[5] Xu Y, Kanauchi A, von Arnim AG, Piston DW, Johnson CH (2003) Bioluminescence resonance energy transfer: monitoring protein-protein interactions in living cells. Methods Enzymol 360:289–301 - DOI - PubMed

[6] Xu Y, Kanauchi A, von Arnim AG, Piston DW, Johnson CH (2003) Bioluminescence resonance energy transfer: monitoring protein-protein interactions in living cells. Methods Enzymol 360:289–301 - DOI - PubMed

[7] Wu PG, Brand L (1994) Resonance energy transfer: methods and applications. Anal Biochem 218:1–13 - DOI - PubMed

[8] Wu PG, Brand L (1994) Resonance energy transfer: methods and applications. Anal Biochem 218:1–13 - DOI - PubMed

[9] Siddiqui S, Cong W-N, Daimon CM, Martin B, Maudsley S (2013) BRET biosensor analysis of receptor tyrosine kinase functionality. Front Endocrinol (Lausanne) 4:46 - DOI - PubMed

[10] Siddiqui S, Cong W-N, Daimon CM, Martin B, Maudsley S (2013) BRET biosensor analysis of receptor tyrosine kinase functionality. Front Endocrinol (Lausanne) 4:46 - DOI - PubMed

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Measuring NLR Oligomerization III: Detection of NLRP3 and NLRC4 Complex by Bioluminescence Resonance Energy Transfer

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