A fluorogenic probe for SNAP-tagged plasma membrane proteins based on the solvatochromic molecule Nile Red.
A fluorogenic probe for plasma membrane proteins based on the dye Nile Red and SNAP-tag is introduced. It takes advantage of Nile Red, a solvatochromic molecule highly fluorescent in an apolar environment, such as cellular membranes, but almost dark in a polar aqueous environment. The probe possesses a tuned affinity for membranes allowing its Nile Red moiety to insert into the lipid bilayer of the plasma membrane, becoming fluorescent, only after its conjugation to a SNAP-tagged plasma membrane protein. The fluorogenic character of the probe was demonstrated for different SNAP-tag fusion proteins, including the human insulin receptor. This work introduces a new approach for generating a powerful turn-on probe for "no-wash" labeling of plasma membrane proteins with numerous applications in bioimaging.
École Polytechnique Fédérale de Lausanne (EPFL) , Institute of Chemical Sciences and Engineering, National Centre of Competence in Research (NCCR) in Chemical Biology, 1015 Lausanne, Switzerland.
Efthymia
E
Prifti
Luc
L
Reymond
Miwa
M
Umebayashi
Ruud
R
Hovius
Howard
H
Riezman
Kai
K
Johnsson
D000818:Animals; D015703:Antigens, CD / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016466:CHO Cells; D002256:Carbonic Anhydrases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D004579:Electron Transport; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D008051:Lipid Bilayers / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D057054:Molecular Imaging / Q000379:methods*; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010078:Oxazines / Q000737:chemistry*; D011972:Receptor, Insulin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D015703:Antigens, CD; D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D008565:Membrane Proteins; D010078:Oxazines; D011993:Recombinant Fusion Proteins; D019853:O(6)-Methylguanine-DNA Methyltransferase; C435941:INSR protein, human; D011972:Receptor, Insulin; D002256:Carbonic Anhydrases; C044808:nile red