A microscope-based screening platform for large-scale functional protein analysis in intact cells.
A modular microscope-based screening platform, with applications in large-scale analysis of protein function in intact cells is described. It includes automated sample preparation, image acquisition, data management and analysis, and the genome-wide automated retrieval of bioinformatic information. The modular nature of the system ensures that it is rapidly adaptable to new biological questions or sets of proteins. Two automated functional assays addressing protein secretion and the integrity of the Golgi complex were developed and tested. This shows the potential of the system in large-scale, cell-based functional proteomic projects.
Liebel
Urban
U
Cell Biology and Cell Biophysics Programme, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg, Germany.
Starkuviene
Vytaute
V
Erfle
Holger
H
Simpson
Jeremy C
JC
Poustka
Annemarie
A
Wiemann
Stefan
S
Pepperkok
Rainer
R
D001331:Automation / Q000295:instrumentation / Q000379:methods; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015894:Genome, Human; D006056:Golgi Apparatus / Q000502:physiology / Q000648:ultrastructure; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D010455:Peptides / Q000032:analysis* / Q000378:metabolism; D021381:Protein Transport; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods; D013223:Statistics as Topic / Q000379:methods; D014162:Transfection / Q000379:methods
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D010455:Peptides; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins