Mapping the dynamics and nanoscale organization of synaptic adhesion proteins using monomeric streptavidin.
Nat Comms. 2016-03-16; 7: 10773
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1. Nat Commun. 2016 Mar 16;7:10773. doi: 10.1038/ncomms10773.
Mapping the dynamics and nanoscale organization of synaptic adhesion proteins
using monomeric streptavidin.
Chamma I(1)(2), Letellier M(1)(2), Butler C(1)(2)(3), Tessier B(1)(2), Lim KH(4),
Gauthereau I(1)(2), Choquet D(1)(2)(5), Sibarita JB(1)(2), Park S(4), Sainlos
M(1)(2), Thoumine O(1)(2).
(1)Interdisciplinary Institute for Neuroscience, UMR 5297, Centre National de la
Recherche Scientifique, 33077 Bordeaux, France.
(2)Interdisciplinary Institute for Neuroscience, University of Bordeaux, 33077
(3)Imagine Optic, 18 rue Charles de Gaulle, 91400 Orsay, France.
(4)Department of Chemical and Biological Engineering, University at Buffalo,
Buffalo, New York 14260, USA.
(5)Bordeaux Imaging Center, UMS 3420 Centre National de la Recherche
Scientifique, University of Bordeaux, US 4 INSERM, 33077 Bordeaux, France.
The advent of super-resolution imaging (SRI) has created a need for optimized
labelling strategies. We present a new method relying on fluorophore-conjugated
monomeric streptavidin (mSA) to label membrane proteins carrying a short,
enzymatically biotinylated tag, compatible with SRI techniques including uPAINT,
STED and dSTORM. We demonstrate efficient and specific labelling of target
proteins in confined intercellular and organotypic tissues, with reduced steric
hindrance and no crosslinking compared with multivalent probes. We use mSA to
decipher the dynamics and nanoscale organization of the synaptic adhesion
molecules neurexin-1β, neuroligin-1 (Nlg1) and leucine-rich-repeat transmembrane
protein 2 (LRRTM2) in a dual-colour configuration with GFP nanobody, and show
that these proteins are diffusionally trapped at synapses where they form apposed
trans-synaptic adhesive structures. Furthermore, Nlg1 is dynamic, disperse and
sensitive to synaptic stimulation, whereas LRRTM2 is organized in compact and
stable nanodomains. Thus, mSA is a versatile tool to image membrane proteins at
high resolution in complex live environments, providing novel information about
the nano-organization of biological structures.
PMID: 26979420 [Indexed for MEDLINE]