Evaluation of metabolic labeling for comparative proteomics in breast cancer cells.
Protein expression patterns in the cytosol of MCF-7 cells resistant to adriamycin and to adriamycin/verapamil were compared to that of the parental MCF-7 cell line and to each other using metabolic labeling and two-dimensional gel electrophoresis. Growing the parental MCF-7 cell line in 13C6-arginine- and 13C6-lysine-enriched medium resulted in C-terminal labeling of all tryptic peptides. The culture media was optimized for the incorporation of these labeled amino acids under conditions that also supported cell growth. Protein abundances were found to be distinctive in MCF-7 cells resistant to adriamycin and those selected for resistance to both adriamycin and verapamil.
Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA.
Marion L
ML
Gehrmann
Yetrib
Y
Hathout
Catherine
C
Fenselau
D017302:Annexins / Q000032:analysis / Q000378:metabolism; D001120:Arginine / Q000737:chemistry / Q000378:metabolism; D001943:Breast Neoplasms / Q000737:chemistry / Q000378:metabolism / Q000473:pathology; D002247:Carbon Isotopes; D045744:Cell Line, Tumor; D003903:Deuterium; D004317:Doxorubicin / Q000494:pharmacology; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D005954:Glucosephosphate Dehydrogenase / Q000032:analysis / Q000378:metabolism; D005982:Glutathione Transferase / Q000032:analysis / Q000378:metabolism; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D008239:Lysine / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry; D010544:Peroxidases / Q000032:analysis / Q000378:metabolism; D054464:Peroxiredoxins; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D014357:Trypsin / Q000378:metabolism; D043222:Ubiquitin Thiolesterase / Q000032:analysis / Q000378:metabolism; D014700:Verapamil / Q000494:pharmacology
D017302:Annexins; D002247:Carbon Isotopes; D011506:Proteins; C482860:UCHL1 protein, human; D004317:Doxorubicin; D001120:Arginine; D003903:Deuterium; D014700:Verapamil; D005954:Glucosephosphate Dehydrogenase; D010544:Peroxidases; D054464:Peroxiredoxins; D005982:Glutathione Transferase; D043222:Ubiquitin Thiolesterase; D014357:Trypsin; D008239:Lysine