High-throughput generation of selected reaction-monitoring assays for proteins and proteomes.
Selected reaction monitoring (SRM) uses sensitive and specific mass spectrometric assays to measure target analytes across multiple samples, but it has not been broadly applied in proteomics owing to the tedious assay development process for each protein. We describe a method based on crude synthetic peptide libraries for the high-throughput development of SRM assays. We illustrate the power of the approach by generating and applying validated SRM assays for all Saccharomyces cerevisiae kinases and phosphatases.
Picotti
Paola
P
Institute of Molecular Systems Biology, Swiss Federal Institute of Technology, Zurich, Switzerland.
Rinner
Oliver
O
Stallmach
Robert
R
Dautel
Franziska
F
Farrah
Terry
T
Domon
Bruno
B
Wenschuh
Holger
H
Aebersold
Ruedi
R
D001681:Biological Assay / Q000379:methods*; D030562:Databases, Protein; D057166:High-Throughput Screening Assays / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D019151:Peptide Library; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism; D011506:Proteins / Q000032:analysis*; D020543:Proteome / Q000032:analysis*; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000201:enzymology
D019151:Peptide Library; D011506:Proteins; D020543:Proteome; D011494:Protein Kinases; D010744:Phosphoric Monoester Hydrolases