title: High-resolution subunit detection of glutamate receptor by ultrasmall gold nanoparticles.
abstract: In this study, we aimed to increase the sensitivity of protein labeling using 1.4 nm gold nanoparticles and glutamate δ2 receptor (GluD2) from the postsynaptic membrane of the Purkinje cells. The very small marker size of the particles reduces the steric hindrance between antibodies leading to a higher labeling efficiency of more than one subunit per single receptor molecule. The nanoparticles are visible in 200 kV dark-field scanning transmission electron microscope on freeze-fractured carbon replica of nervous tissue after plasma cleaning treatment. The different elemental composition of nanoparticles as Au nanogold or CdS quantum dot can be distinguished by energy dispersive X-ray spectroscopy. This method ensures detection of an average of three subunits per GluD2 and often labels all four of them with 1.4 nm Au nanoparticles. It is concluded that this high-resolution microscopic method is useful for exploring the quaternary structure of membrane proteins.
authors.affiliation: Laboratory of Engineering Nanobiotechnology, Department of Engineering Geoecology, University of Mining and Geology "St. Ivan Rilski," Stoyan Edrev Str., Sofia 1700, Bulgaria. alexandre_loukanov@abv.bg
authors.forename: Alexandre R
authors.initials: AR
authors.lastname: Loukanov
authors.forename: Hristo
authors.initials: H
authors.lastname: Gagov
mesh_terms: D000818:Animals; D020285:Cryoelectron Microscopy; D006046:Gold / Q000032:analysis* / Q000378:metabolism*; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles; D021122:Protein Subunits / Q000032:analysis; D011689:Purkinje Cells / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017470:Receptors, Glutamate / Q000032:analysis*; D013052:Spectrometry, X-Ray Emission; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D021122:Protein Subunits; D017470:Receptors, Glutamate; D006046:Gold