Targeted comparative proteomics employing nano liquid chromatography coupled with Fourier transform ion cyclotron resonance mass spectrometry (nLC-FTICR MS)

ANYL 294

Hyuk Kang, hyuk.kang@pnl.gov, Michael A. Buschbach, Navdeep Jaitly, Nikola Tolic, David C. Prior, Mary S Lipton, Ljiljana Pasa-Tolic, and Richard D. Smith, rds@pnl.gov. Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99352
A targeted tandem mass spectrometry (i.e., MS/MS), an approach suitable for characterization of peptides showing statistically significant changes in abundance after a perturbation, will be illustrated using 14N/15N metabolic labeling to evaluate the effect of O2 limitation on Shewanella oneidensis growth. An attention list, specifying mass-to-charge ratios and LC elution times of differentially abundant peptides, was generated from nLC-FTICR MS-only data. Interesting peptides were subsequently identified employing targeted MS/MS in a hybrid FTICR spectrometer equipped with external ion selection, accumulation, and fragmentation capabilities. In this experiment the attention list directed the MS/MS acquisition to specifically target the species having the greatest potential for being markers of a specific state, using software and hardware developed in-house. Accurate selection of targets, which proved to be critical for effective analysis of a complex peptide mixture, was enabled by high mass resolution and accuracy offered by FTICR MS and efficient on-the-fly LC-elution time correction.
 

Analytical Approaches: Mass Spectrometry
1:30 PM-4:20 PM, Wednesday, 13 September 2006 Moscone Center -- Room 124, Oral

Division of Analytical Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006