Semisynthetic fluorescent pH sensors for imaging exocytosis and endocytosis. Issue 1 (December 2017)

• Record Type: Journal Article
• Authors: Martineau, Magalie; Somasundaram, Agila; Grimm, Jonathan; Gruber, Todd; Choquet, Daniel; Taraska, Justin; Lavis, Luke; Perrais, David
• Is Part Of: Nature communications. Volume 8:Issue 1(2017)
• Page Start: 1
• Page End: 10
• Publication Date: 2017-12
• Subjects: Biology -- Periodicals; Physical sciences -- Periodicals; 505
• DOI: 10.1038/s41467-017-01752-5
• Languages: English
• ISSNs: 2041-1723

Abstract

Abstract The GFP-based superecliptic pHluorin (SEP) enables detection of exocytosis and endocytosis, but its performance has not been duplicated in red fluorescent protein scaffolds. Here we describe "semisynthetic" pH-sensitive protein conjugates with organic fluorophores, carbofluorescein, and Virginia Orange that match the properties of SEP. Conjugation to genetically encoded self-labeling tags or antibodies allows visualization of both exocytosis and endocytosis, constituting new bright sensors for these key steps of synaptic transmission. Existing pH-sensitive red fluorescent protein probes don't perform well in monitoring exocytosis and endocytosis. Here, the authors combine organic dyes with self-labeling tags or antibodies to develop semisynthetic protein conjugates that can image synaptic vesicle fusion events in living cells.