Integration of 18O labeling and solution isoelectric focusing in a shotgun analysis of mitochondrial proteins.
Forward and reverse (18)O labeling are integrated with solution isoelectric focusing and capillary LC-tandem mass spectrometry to evaluate a new strategy for quantitative proteomics and to study abundance changes in mitochondrial proteins associated with drug resistance in MCF-7 human cancer cells. Galectin-3 binding protein, which is involved in apoptosis, was detected only in the resistant cell line, as a result of reverse labeling. Among 278 proteins identified, 12 were detected with abundances altered at least 2-fold.
Wang
Jinshan
J
Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA.
Gutierrez
Peter
P
Edwards
Nathan
N
Fenselau
Catherine
C
D045744:Cell Line, Tumor; D005591:Chemical Fractionation; D019008:Drug Resistance, Neoplasm; D006801:Humans; D007525:Isoelectric Focusing; D024101:Mitochondrial Proteins / Q000032:analysis* / Q000737:chemistry*; D008942:Mitoxantrone / Q000494:pharmacology; D009842:Oligopeptides / Q000032:analysis / Q000737:chemistry; D010103:Oxygen Isotopes / Q000378:metabolism; D012996:Solutions; D013194:Staining and Labeling; D053719:Tandem Mass Spectrometry
D024101:Mitochondrial Proteins; D009842:Oligopeptides; D010103:Oxygen Isotopes; D012996:Solutions; D008942:Mitoxantrone