A three-symbol code for organized proteomes based on cyclical imaging of protein locations.
BACKGROUND
A major challenge in the post genomic era is to map and decipher the functional molecular networks of proteins directly in a cell or a tissue. This task requires technologies for the colocalization of random numbers of different molecular components (e.g. proteins) in one sample in one experiment.


METHODS
Multi-epitope-ligand-"kartographie" (MELK) was developed as a microscopic imaging technology running cycles of iterative fluorescence tagging, imaging, and bleaching, to colocalize a large number of proteins in one sample (morphologically intact routinely fixed cells or tissue).


RESULTS
In the present study, 18 different cell surface proteins were colocalized by MELK in cells and tissue sections in different compartments of the human immune system. From the resulting sets of multidimensional binary vectors the most prominent groups of protein-epitope arrangements were extracted and imaged as protein "toponome" maps providing direct insight in the higher order topological organization of immune compartments uncovering new tissue domains. The data sets suggest that protein networks, topologically organized in proteomes in situ, obey a unique protein-colocation and -anticolocation code describable by three symbols.


CONCLUSIONS
The technology has the potential to colocalize hundreds of proteins and other molecular components in one sample and may offer many applications in biology and medicine.
Schubert
Walter
W
Molecular Pattern Recognition Research Group, Institute of Medical Neurobiology, University of Magdeburg, Germany. walter.schubert@medizin.uni-magdeburg.de
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000032:analysis* / Q000276:immunology; D006680:HLA Antigens / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D009132:Muscles / Q000276:immunology; D014066:Palatine Tonsil / Q000276:immunology; D040081:Protein Array Analysis; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D006680:HLA Antigens