Takashi Kanadome, Natsumi Hoshino, Takeharu Nagai, Tomoki Matsuda, Takeshi Yagi (2021) Development of FRET-based indicators for visualizing homophilic trans interaction of a clustered protocadherin., Scientific reports, Volume 11, Number 1, pp. 22237
Published in 2021 Nov 15 (Electronic publication in Nov. 15, 2021, midnight )
(Abstract) Clustered protocadherins (Pcdhs), which are cell adhesion molecules, play a fundamental role in self-recognition and non-self-discrimination by conferring diversity on the cell surface. Although systematic cell-based aggregation assays provide information regarding the binding properties of Pcdhs, direct visualization of Pcdh trans interactions across cells remains challenging. Here, we present Forster resonance energy transfer (FRET)-based indicators for directly visualizing Pcdh trans interactions. We developed the indicators by individually inserting FRET donor and acceptor fluorescent proteins (FPs) into the ectodomain of Pcdh molecules. They enabled successful visualization of specific trans interactions of Pcdh and revealed that the Pcdh trans interaction is highly sensitive to changes in extracellular Ca(2+) levels. We expect that FRET-based indicators for visualizing Pcdh trans interactions will provide a new approach for investigating the roles of Pcdh in self-recognition and non-self-discrimination processes.(MeSH Terms)