Measurement of Protein-Protein Interactions ...
Type de document :
Compte-rendu et recension critique d'ouvrage
DOI :
PMID :
Titre :
Measurement of Protein-Protein Interactions through Microscale Thermophoresis (MST)
Auteur(s) :
Romain, Magnez [Auteur]
Cancer Heterogeneity, Plasticity and Resistance to Therapies - UMR 9020 - U 1277 [CANTHER]
Thiroux, Bryan [Auteur]
Cancer Heterogeneity, Plasticity and Resistance to Therapies - UMR 9020 - U 1277 [CANTHER]
Tardy, Morgane [Auteur]
Cancer Heterogeneity, Plasticity and Resistance to Therapies - UMR 9020 - U 1277 [CANTHER]
Quesnel, Bruno [Auteur]
Cancer Heterogeneity, Plasticity and Resistance to Therapies - UMR 9020 - U 1277 [CANTHER]
Thuru, Xavier [Auteur correspondant]
Cancer Heterogeneity, Plasticity and Resistance to Therapies - UMR 9020 - U 1277 [CANTHER]
Cancer Heterogeneity, Plasticity and Resistance to Therapies - UMR 9020 - U 1277 [CANTHER]
Thiroux, Bryan [Auteur]
Cancer Heterogeneity, Plasticity and Resistance to Therapies - UMR 9020 - U 1277 [CANTHER]
Tardy, Morgane [Auteur]
Cancer Heterogeneity, Plasticity and Resistance to Therapies - UMR 9020 - U 1277 [CANTHER]
Quesnel, Bruno [Auteur]
Cancer Heterogeneity, Plasticity and Resistance to Therapies - UMR 9020 - U 1277 [CANTHER]
Thuru, Xavier [Auteur correspondant]
Cancer Heterogeneity, Plasticity and Resistance to Therapies - UMR 9020 - U 1277 [CANTHER]
Titre de la revue :
Bio-protocol
Éditeur :
Bio-protocol LCC
Date de publication :
2020-04-05
ISSN :
2331-8325
Mot(s)-clé(s) en anglais :
Microscale thermophoresis
Immune escape
PD-L1
PD-1
Protein-protein interaction (PPI)
Screening
Cell lysate
Purification-free protocol
Immune escape
PD-L1
PD-1
Protein-protein interaction (PPI)
Screening
Cell lysate
Purification-free protocol
Discipline(s) HAL :
Sciences du Vivant [q-bio]
Résumé en anglais : [en]
The binding interactions of PD-1 and PD-L1 have been studied by surface plasmonresonance (SPR) and isothermal titration calorimetry (ITC) over the past few years, but theseinvestigations resulted in controversy regarding ...
Lire la suite >The binding interactions of PD-1 and PD-L1 have been studied by surface plasmonresonance (SPR) and isothermal titration calorimetry (ITC) over the past few years, but theseinvestigations resulted in controversy regarding the values of the dissociation constant (Kd) (Freeman et al., 2000). MST is a powerful new method for the quantitative analysis of protein-protein interactions (PPIs) with low sample consumption. The technique is based on the movement of molecules along microscopic temperature gradients, and it detects changes in their hydration shell, charge or size. One binding partner is fluorescently labeled, while the other binding partner remains label-free. We used a protocol that allows the determination of the binding affinity by MST without purification of the target protein from the cell lysate. The application of this MST method to PD-1-eGFP and PD-L1-eGFP expressed in CHO-K1 cells allowed us, for the first time, to determine the affinity of the complex formed between PD-1 and its ligand PD-L1 during tumor escape. The protocol has a variety of potential applications for studying the interactions of proteins with small molecules.Lire moins >
Lire la suite >The binding interactions of PD-1 and PD-L1 have been studied by surface plasmonresonance (SPR) and isothermal titration calorimetry (ITC) over the past few years, but theseinvestigations resulted in controversy regarding the values of the dissociation constant (Kd) (Freeman et al., 2000). MST is a powerful new method for the quantitative analysis of protein-protein interactions (PPIs) with low sample consumption. The technique is based on the movement of molecules along microscopic temperature gradients, and it detects changes in their hydration shell, charge or size. One binding partner is fluorescently labeled, while the other binding partner remains label-free. We used a protocol that allows the determination of the binding affinity by MST without purification of the target protein from the cell lysate. The application of this MST method to PD-1-eGFP and PD-L1-eGFP expressed in CHO-K1 cells allowed us, for the first time, to determine the affinity of the complex formed between PD-1 and its ligand PD-L1 during tumor escape. The protocol has a variety of potential applications for studying the interactions of proteins with small molecules.Lire moins >
Langue :
Français
Vulgarisation :
Non
Collections :
Source :
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