title: Spatial distribution and structural arrangement of a murine cytomegalovirus glycoprotein detected by SPDM localization microscopy.
abstract: Novel approaches of localization microscopy have opened new insights into the molecular nano-cosmos of cells. We applied a special embodiment called spectral position determination microscopy (SPDM) that has the advantage to run with standard fluorescent dyes or proteins under standard preparation conditions. Pointillist images with a resolution in the order of 10 nm can be obtained by SPDM. Therefore, vector pEYFP-m164, encoding the murine cytomegalovirus glycoprotein gp36.5/m164 fused to enhanced yellow fluorescent protein, was transiently transfected into COS-7 cells. This protein shows exceptional intracellular trafficking dynamics, moving within the endoplasmic reticulum (ER) and outer nuclear membrane. The molecular positions of gp36.5/m164 were visualized and determined by SPDM imaging. From the position point patterns of the protein molecules, their arrangements were quantified by next neighbour distance analyses. Three different structural arrangements were discriminated: (a) a linear distribution along the membrane, (b) a highly structured distribution in the ER, and (c) a homogenous distribution in the cellular cytoplasm. The results indicate that the analysis of next neighbour distances on the nano-scale allows the identification and discrimination of different structural arrangements of molecules within their natural cellular environment.
authors: Kirchhoff-Institute for Physics, Im Neuenheimer Feld 227, 69120, Heidelberg, Germany.
authors: Patrick
authors: P
authors: Müller
authors: Niels A
authors: NA
authors: Lemmermann
authors: Rainer
authors: R
authors: Kaufmann
authors: Manuel
authors: M
authors: Gunkel
authors: Daniel
authors: D
authors: Paech
authors: Georg
authors: G
authors: Hildenbrand
authors: Rafaela
authors: R
authors: Holtappels
authors: Christoph
authors: C
authors: Cremer
authors: Michael
authors: M
authors: Hausmann
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry; D019556:COS Cells; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D006023:Glycoproteins / Q000032:analysis* / Q000235:genetics; D008164:Luminescent Proteins / Q000737:chemistry; D051379:Mice; D008856:Microscopy, Fluorescence; D018146:Muromegalovirus / Q000737:chemistry*; D014759:Viral Envelope Proteins / Q000032:analysis* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D006023:Glycoproteins; D008164:Luminescent Proteins; D014759:Viral Envelope Proteins; C054562:yellow fluorescent protein, Bacteria