Authors
Junmin Peng, Joshua E Elias, Carson C Thoreen, Larry J Licklider, Steven P Gygi
Publication date
2003/2/3
Journal
Journal of proteome research
Volume
2
Issue
1
Pages
43-50
Publisher
American Chemical Society
Description
Highly complex protein mixtures can be directly analyzed after proteolysis by liquid chromatography coupled with tandem mass spectrometry (LC−MS/MS). In this paper, we have utilized the combination of strong cation exchange (SCX) and reversed-phase (RP) chromatography to achieve two-dimensional separation prior to MS/MS. One milligram of whole yeast protein was proteolyzed and separated by SCX chromatography (2.1 mm i.d.) with fraction collection every minute during an 80-min elution. Eighty fractions were reduced in volume and then re-injected via an autosampler in an automated fashion using a vented-column (100 μm i.d.) approach for RP-LC−MS/MS analysis. More than 162 000 MS/MS spectra were collected with 26 815 matched to yeast peptides (7537 unique peptides). A total of 1504 yeast proteins were unambiguously identified in this single analysis. We present a comparison of this …
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Scholar articles
J Peng, JE Elias, CC Thoreen, LJ Licklider, SP Gygi - Journal of proteome research, 2003