title: Targeted glycoproteomic identification of biomarkers for human breast carcinoma.
abstract: Glycosylation is a dynamic post-translational modification that changes during the development and progression of various malignancies. During the oncogenesis of breast carcinoma, the glycosyltransferase known as N-acetylglucosaminyltransferase Va (GnT-Va) transcript levels and activity are increased due to activated oncogenic signaling pathways. Elevated GnT-V levels leads to increased beta(1,6)-branched N-linked glycan structures on glycoproteins that can be measured using a specific carbohydrate binding protein or lectin known as L-PHA. L-PHA does not bind to nondiseased breast epithelial cells, but during the progression to invasive carcinoma, cells show a progressive increase in L-PHA binding. We have developed a procedure for intact protein L-PHA-affinity enrichment, followed by nanospray ionization mass spectrometry (NSI-MS/MS), to identify potential biomarkers for breast carcinoma. We identified L-PHA reactive glycoproteins from matched normal (nondiseased) and malignant tissue isolated from patients with invasive ductal breast carcinoma. Comparison analysis of the data identified 34 proteins that were enriched by L-PHA fractionation in tumor relative to normal tissue for at least 2 cases of ductal invasive breast carcinoma. Of these 34 L-PHA tumor enriched proteins, 12 are common to all 4 matched cases analyzed. These results indicate that lectin enrichment strategies targeting a particular glycan change associated with malignancy can be an effective method of identifying potential biomarkers for breast carcinoma.
authors.affiliation: Complex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia 30605, USA. kabbott@uga.edu
authors.forename: Karen L
authors.initials: KL
authors.lastname: Abbott
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Aoki
authors.forename: Jae-Min
authors.initials: JM
authors.lastname: Lim
authors.forename: Mindy
authors.initials: M
authors.lastname: Porterfield
authors.forename: Rachelle
authors.initials: R
authors.lastname: Johnson
authors.forename: Ruth M
authors.initials: RM
authors.lastname: O'Regan
authors.forename: Lance
authors.initials: L
authors.lastname: Wells
authors.forename: Michael
authors.initials: M
authors.lastname: Tiemeyer
authors.forename: Michael
authors.initials: M
authors.lastname: Pierce
mesh_terms: D000951:Antigens, Neoplasm / Q000032:analysis; D014408:Biomarkers, Tumor / Q000032:analysis*; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D015815:Cell Adhesion Molecules / Q000032:analysis; D016326:Extracellular Matrix Proteins / Q000032:analysis; D005260:Female; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry; D006242:Haptoglobins / Q000032:analysis; D006801:Humans; D025942:Oligosaccharides, Branched-Chain / Q000032:analysis; D010835:Phytohemagglutinins / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D015854:Up-Regulation
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D015815:Cell Adhesion Molecules; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; C058632:HPR protein, human; D006242:Haptoglobins; D025942:Oligosaccharides, Branched-Chain; C512835:POSTN protein, human; D010835:Phytohemagglutinins