A Phosphoproteomic Study Of Insulin Signaling Pathway Using A Novel High-Throughput Pipeline

Аннотация

Recent advances in the speed and sensitivity of mass spectrometers and analytical methods, the exponential acceleration of computer processing speeds, and the availability of genomic databases from an array of species and protein information databases have led to a deluge of proteomic data. Unfortunately, this enhancement in data acquisition has not been accompanied by a concomitant increase in the availability of tools allowing users to rapidly assimilate, explore, and analyze this data and adapt to a variety of experimental workflows with minimal user intervention. Often the manual aggregation and analysis of proteomic data in current proteomics software distract investigators from the biological meaning of their data, leading to the all-too-frequent deposition of proteomic data into the scientific literature with little or no biological or clinical interpretation. We seek to fill the gap by providing a flexible platform for high-throughput autonomous proteomic analysis with the following critical components: liquid chromatography/mass spectrometry (LC/MS) acquisition control module, tandem mass spectra (MS/MS) database search, peptide spectral validation, peptide quantitation, quantitative data exploration tool within a relational database, cached public protein information databases and protein network exploration tool. The LC/MS control tool integrates lab information management system (LIMS) to provide automated multidimensional sample analysis, as well as captures meta-data during analysis and associates them with sample preparation protocols and experiment results in a relational database. Instrument acquired raw data are streamlined through a customized proteomic pipeline for database searching followed by peptide validation