Analyzing proteome topology and function by automated multidimensional fluorescence microscopy.
Temporal and spatial regulation of proteins contributes to function. We describe a multidimensional microscopic robot technology for high-throughput protein colocalization studies that runs cycles of fluorescence tagging, imaging and bleaching in situ. This technology combines three advances: a fluorescence technique capable of mapping hundreds of different proteins in one tissue section or cell sample; a method selecting the most prominent combinatorial molecular patterns by representing the data as binary vectors; and a system for imaging the distribution of these protein clusters in a so-called toponome map. By analyzing many cell and tissue types, we show that this approach reveals rules of hierarchical protein network organization, in which the frequency distribution of different protein clusters obeys Zipf's law, and state-specific lead proteins appear to control protein network topology and function. The technology may facilitate the development of diagnostics and targeted therapies.
Schubert
Walter
W
Molecular Pattern Recognition Research (MPRR) Group, Institute of Medical Neurobiology, Otto-von-Guericke-University Magdeburg, D-39120 Magdeburg, Germany. walter.schubert@medizin.uni-magdeburg.de
Bonnekoh
Bernd
B
Pommer
Ansgar J
AJ
Philipsen
Lars
L
Böckelmann
Raik
R
Malykh
Yanina
Y
Gollnick
Harald
H
Friedenberger
Manuela
M
Bode
Marcus
M
Dress
Andreas W M
AW
D003876:Dermatitis, Atopic / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence / Q000379:methods*; D010336:Pathology / Q000379:methods; D011506:Proteins / Q000032:analysis* / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D011565:Psoriasis / Q000378:metabolism; D015203:Reproducibility of Results; D012867:Skin / Q000378:metabolism
D011506:Proteins