New algorithm for 15N/14N quantitation with LC-ESI-MS using an LTQ-FT mass spectrometer.
A new algorithm (QN) for the (15)N /(14)N quantitation of relative protein abundances in complex proteomic samples is described. QN takes advantage of the high resolution, mass accuracy and throughput of the hybrid mass spectrometer LTQ-FT MS. Peptide quantitation is based on MS peak intensity (measured in the FT MS), while peptide identification is performed in the MS/MS mode (measured in the LTQ linear ion trap). Accuracy of the protein abundance is enhanced by a novel scoring procedure, allowing filtering of less reliable measurements of peptide abundances. The performance of QN is illustrated in the relative quantitative analysis of M. acetivorans C2A cultures grown with carbon monoxide vs methanol as substrate. Roughly 1,000 proteins were quantitated with an average CV of 9% for the protein abundance ratios. QN performs quantitation without manual intervention, does not require high processing power, and generates files compatible with the Guidelines for Proteomic Data Publication.
Andreev
Victor P
VP
Barnett Institute and Department of Chemistry, Northeastern University, Boston, Massachusetts 02115, USA.
Li
Lingyun
L
Rejtar
Tomas
T
Li
Qingbo
Q
Ferry
James G
JG
Karger
Barry L
BL
D000465:Algorithms; D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002853:Chromatography, Liquid; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods; D017020:Methanosarcina / Q000737:chemistry; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000032:analysis*; D009587:Nitrogen Isotopes / Q000032:analysis*; D010455:Peptides / Q000032:analysis / Q000235:genetics; D040901:Proteomics / Q000379:methods; D012984:Software
D019843:Archaeal Proteins; D009587:Nitrogen Isotopes; D010455:Peptides; D009584:Nitrogen