Proximity ligation assay to study protein-protein interactions of proteins on two different cells

BioTechniques
Rushikesh SableSeetharama D Jois

Abstract

Protein-protein interactions (PPI) by homo-, hetero- or oligo-merization in the cellular environment regulate cellular processes. PPI can be inhibited by antibodies, small molecules or peptides, and this inhibition has therapeutic value. A recently developed method, the proximity ligation assay (PLA), provides detection of PPI in the cellular environment. However, most applications using this assay are for proteins expressed in the same cell. We employ PLA for the first time to study PPI of cell surface proteins on two different cells. Inhibition of PPI using a peptide inhibitor is also quantified using this assay; PLA is used to detect PPI of CD2 and CD58 between Jurkat cells (T cells) and human fibroblast-like synoviocyte-rheumatoid arthritis cells that are important in the immune response in the autoimmune disease rheumatoid arthritis. This assay provides direct evidence of inhibition of PPI of two proteins on different cell surfaces.

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Citations

Oct 15, 2019·Synapse·Gabriel López-RamírezBenjamín Florán
Apr 3, 2020·Laboratory Investigation; a Journal of Technical Methods and Pathology·Yakun LiuBin Gong
Mar 6, 2021·European Journal of Medicinal Chemistry·Sitanshu S SinghSeetharama Jois
Jul 25, 2019·Bioconjugate Chemistry·Igor DovganAlain Wagner
Aug 24, 2021·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·Yong Wan, Heather L Szabo-Rogers

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Methods Mentioned

BETA
proximity ligation assay
proximity ligation
flow cytometry
confocal microscopy

Software Mentioned

ImageJ
CellQuest Pro
FlowJo
CellSens dimension
SAS

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