Comparative study of the localization and conformation of immunocomplexes single molecule of NMDA receptor protein on neuron membrane by AFM
- VernacularTitle:神经元膜NMDA受体蛋白免疫复合物单分子纳米结构及定位AFM比较研究
- Author:
Yu YI-GANG
1
;
Xu RU-XIANG
;
Jiang XIAO-DAN
;
Cai YING-QIAN
;
Ke YI-QUAN
;
Yao QIAN-MING
Author Information
1. 南方医科大学
- Keywords:
neuron;
NMDA receptor;
membrane protein;
single molecule;
atomic force microscope;
immunocytochemistry
- From:
Neuroscience Bulletin
2005;21(2):117-128
- CountryChina
- Language:Chinese
-
Abstract:
Objective In order to study the localization of NMDA receptor protein on the neuron membrane, a new immunocytochemistry method to visualize and quantify 3D earmark the membrane protein in nanometer scale has been achieved by atomic force microscope (AFM) with labeled colloid gold particle. Methods First step, the distribution of the anti-NMDAR1 IgG-SPA-gold complexes molecule on the surface of mica had been scanned by AFM and the characteristic 3D conformation was analyzed by particle software. Then the 3D conformation was contrasted with the IgG-SPA-gold complexes molecules combined with the neuron membrane surface and that were compared with the results of light microscope, laser confocal microscope and scanning electron microscope. Results The 3D conformation of IgG-SPA-gold scanned through mica surface shows the average diameter of globe particles is 49 nm. The neuron membrane NMDA receptor protein combined with IgG-SPA-gold immunocomplexes are globe or cosh shapes . Its average diameter is 53 nm. The long axis section is a typical two apexes structure. Conclusion AFM could determine the situation and 3D conformation of NMDAR protein on neuron membrane surface in nanometer scale. NMDAR single molecule could combine with one or two colloidal gold particles. Colloidal gold particle scanned by AFM in situ is a new detecting method of immunocytochemistry.
