Hybridization of pulsed-Q dissociation and collision-activated dissociation in linear ion trap mass spectrometer for iTRAQ quantitation.
Coupling of multiplex isobaric tags for relative and absolute quantitation (iTRAQ) to a sensitive linear ion trap (LTQ) mass spectrometer (MS) is a challenging, but highly promising approach for quantitative high-throughput proteomic profiling. Integration of the advantages of pulsed-Q dissociation (PQD) and collision-activated dissociation (CAD) fragmentation methods into a PQD-CAD hybrid mode, together with PQD optimization and data manipulation with a bioinformatics algorithm, resulted in a robust, sensitive and accurate iTRAQ quantitative proteomic workflow. The workflow was superior to the default PQD setting when profiling the proteome of a gastric cancer cell line, SNU5. Taken together, we established an optimized PQD-CAD hybrid workflow in LTQ-MS for iTRAQ quantitative proteomic profiling that may have wide applications in biological and biomedical research.
School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, and Division of Medical Sciences, National Cancer Centre Singapore, 11 Hospital Drive, Singapore 169610.
Tiannan
T
Guo
Chee Sian
CS
Gan
Huoming
H
Zhang
Yi
Y
Zhu
Oi Lian
OL
Kon
Siu Kwan
SK
Sze
D000465:Algorithms; D000818:Animals; D002364:Caseins / Q000378:metabolism; D002417:Cattle; D045744:Cell Line, Tumor; D002645:Chickens; D019295:Computational Biology / Q000379:methods; D045304:Cytochromes c / Q000378:metabolism; D006736:Horses; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D009113:Muramidase / Q000378:metabolism; D009211:Myoglobin / Q000378:metabolism; D010047:Ovalbumin / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013274:Stomach Neoplasms / Q000378:metabolism / Q000473:pathology
D002364:Caseins; D009211:Myoglobin; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; D045304:Cytochromes c; C421864:hen egg lysozyme; D009113:Muramidase