8C5H

NbSyt1 anti-(rat Synaptotagmin-1) nanobody bound to target cytosolic domain of Synaptotagmin-1

Summary for 8C5H

• Entry DOI: 10.2210/pdb8c5h/pdb
• Descriptor: Synaptotagmin-1, NbSyt1 nanobody, CALCIUM ION, ... (5 entities in total)
• Functional Keywords: nanobody, synaptotagmin 1, metal binding protein
• Biological source: Rattus norvegicus (Norway rat); More
• Total number of polymer chains: 2
• Total formula weight: 29496.10

Authors

Martinez-Carranza, M.,Stenmark, P. (deposition date: 2023-01-09, release date: 2023-08-02, Last modification date: 2024-10-23)

Primary citation

Queiroz Zetune Villa Real, K.,Mougios, N.,Rehm, R.,Sograte-Idrissi, S.,Albert, L.,Rahimi, A.M.,Maidorn, M.,Hentze, J.,Martinez-Carranza, M.,Hosseini, H.,Saal, K.A.,Oleksiievets, N.,Prigge, M.,Tsukanov, R.,Stenmark, P.,Fornasiero, E.F.,Opazo, F.
A Versatile Synaptotagmin-1 Nanobody Provides Perturbation-Free Live Synaptic Imaging And Low Linkage-Error in Super-Resolution Microscopy.
Small Methods, 7:e2300218-e2300218, 2023

PubMed Abstract: Imaging of living synapses has relied for over two decades on the overexpression of synaptic proteins fused to fluorescent reporters. This strategy alters the stoichiometry of synaptic components and ultimately affects synapse physiology. To overcome these limitations, here a nanobody is presented that binds the calcium sensor synaptotagmin-1 (NbSyt1). This nanobody functions as an intrabody (iNbSyt1) in living neurons and is minimally invasive, leaving synaptic transmission almost unaffected, as suggested by the crystal structure of the NbSyt1 bound to Synaptotagmin-1 and by the physiological data. Its single-domain nature enables the generation of protein-based fluorescent reporters, as showcased here by measuring spatially localized presynaptic Ca with a NbSyt1- jGCaMP8 chimera. Moreover, the small size of NbSyt1 makes it ideal for various super-resolution imaging methods. Overall, NbSyt1 is a versatile binder that will enable imaging in cellular and molecular neuroscience with unprecedented precision across multiple spatiotemporal scales.

PubMed: 37421204
DOI: 10.1002/smtd.202300218

Experimental method

X-RAY DIFFRACTION (1.68 Å)