title: HILIC and ERLIC Enrichment of Glycopeptides Derived from Breast and Brain Cancer Cells.
abstract: Aberrant glycosylation has been linked to many different cancer types. The blood-brain barrier (BBB) is a region of the brain that regulates the entrance of ions, diseases, toxins, and so on. However, in breast cancer metastasis, the BBB fails to prevent the crossing of the cancer cells into the brain. Here we present a study of identifying and quantifying the glycosylation of six breast and brain cancer cell lines using hydrophilic interaction liquid chromatography (HILIC) and electrostatic repulsion liquid chromatography (ERLIC) enrichments and LC-MS/MS analysis. Qualitative and quantitative analyses of N-linked glycosylation were performed by both enrichment techniques for individual and complementary comparison. Potential cancer glycopeptide biomarkers were identified and confirmed by chemometric and statistical evaluations. A total of 497 glycopeptides were characterized, of which 401 were common glycopeptides (80.6% overlap) identified from both enrichment techniques. HILIC enrichment yielded 320 statistically significant glycopeptides in 231BR relative to the other cell lines out of 494 unique glycopeptides, and sequential HILIC-ERLIC enrichment yielded 214 statistically significant glycopeptides in 231BR compared with the other cell lines out of 404 unique glycopeptides. The results provide the first comprehensive glycopeptide listing for these six cell lines.
authors.affiliation: Department of Chemistry and Biochemistry, Texas Tech University , Lubbock, Texas 79409, United States.
authors.forename: Lauren G
authors.initials: LG
authors.lastname: Zacharias
authors.affiliation: Department of Chemistry and Biochemistry, Texas Tech University , Lubbock, Texas 79409, United States.
authors.forename: Alyssa K
authors.initials: AK
authors.lastname: Hartmann
authors.affiliation: Department of Chemistry and Biochemistry, Texas Tech University , Lubbock, Texas 79409, United States.
authors.forename: Ehwang
authors.initials: E
authors.lastname: Song
authors.affiliation: Department of Chemistry and Biochemistry, Texas Tech University , Lubbock, Texas 79409, United States.
authors.forename: Jingfu
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Department of Chemistry and Biochemistry, Texas Tech University , Lubbock, Texas 79409, United States.
authors.forename: Rui
authors.initials: R
authors.lastname: Zhu
authors.affiliation: Department of Chemistry and Biochemistry, Texas Tech University , Lubbock, Texas 79409, United States.
authors.forename: Parvin
authors.initials: P
authors.lastname: Mirzaei
authors.affiliation: Department of Chemistry and Biochemistry, Texas Tech University , Lubbock, Texas 79409, United States.
authors.forename: Yehia
authors.initials: Y
authors.lastname: Mechref
mesh_terms: D001932:Brain Neoplasms / Q000737:chemistry* / Q000473:pathology; D001943:Breast Neoplasms / Q000737:chemistry* / Q000473:pathology; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid / Q000379:methods; D003627:Data Interpretation, Statistical; D005260:Female; D006020:Glycopeptides / Q000032:analysis*; D006031:Glycosylation; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008297:Male; D009363:Neoplasm Proteins / Q000032:analysis*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D006020:Glycopeptides; D009363:Neoplasm Proteins
keywords: ERLIC enrichment; HILIC enrichment; N-linked glycosylation; blood−brain barrier; cancer cell lines; glycopeptide; glycoproteomics; mass spectrometry