Uncovering immobilized trypsin digestion features from large-scale proteome data generated by high-resolution mass spectrometry.

ABSTRACT: Immobilized trypsin produces very fast protein digestion, which is attractive for application to high throughput bottom-up proteomics. While there is a rich literature on the preparation of immobilized trypsin, there are very few studies that investigate its application to complex proteomic samples. In this work, we compared solution-phase trypsin with trypsin immobilized on magnetic microspheres for digestion of two complex proteomes, Escherichia coli and the MCF7 cell line. The digests were separated by HPLC, and detected with a Q-Exactive mass spectrometer, which generated high resolution and high quality parent- and fragment-ion mass spectra. The data were analyzed using MaxQuant. We make several conclusions about the features of immobilized trypsin digestion of complex proteomes. Firs

SUBMITTER: Sun L 

PROVIDER: S-EPMC4000775 | biostudies-literature | 2014 Apr

REPOSITORIES: biostudies-literature