Characterization of cancer associated mucin type O-glycans using the exchange sialylation properties of mammalian sialyltransferase ST3Gal-II.

Abstract

Our previous studies suggest that the α2,3sialylated T-antigen (NeuAcα2,3Galβ1,3GalNac-) and associated glycan structures are likely to be elevated during cancer. An easy and reliable strategy to label mucinous glycans that contain such carbohydrates can enable the identification of novel glycoproteins that are cancer associated. To this end, the present study demonstrates that the exchange sialylation property of mammalian ST3Gal-II can facilitate the labeling of mucin glycoproteins in cancer cells, tumor specimens, and glycoproteins in cancer sera. Results show that (i) the radiolabeled mucin glycoproteins of each of the cancer cell lines studied (T47D, MCF7, LS180, LNCaP, SKOV3, HL60, DU4475, and HepG2) is distinct either in terms of the specific glycans presented or their relative distribution. While some cell lines like T47D had only one single sialylated O-glycan, others like LS180 and DU4475 contained a complex mixture of mucinous carbohydrates. (ii) [14C]sialyl labeling of primary tumor cells identified a 25-35 kDa mucin glycoprotein unique to pancreatic tumor. Labeled glycoproteins for other cancers had higher molecular weight. (iii) Studies of [14C] sialylated human sera showed larger mucin glycopeptides and >2-fold larger mucin-type chains in human serum compared to [14C]sialyl labeled glycans of fetuin. Overall, the exchange sialylation property of ST3Gal-II provides an efficient avenue to identify mucinous proteins for applications in glycoproteomics and cancer research.

• Authors: E V Chandrasekaran, Jun Xue, Jie Xia, Robert D Locke, Shilpa A Patil, Sriram Neelamegham, Khushi L Matta
• Journal: Journal of proteome research (J Proteome Res) Vol. 11 Issue 4 Pg. 2609-18 (Apr 06 2012) ISSN: 1535-3907 [Electronic] United States
• PMID: 22329400 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Biomarkers, Tumor
• Carbon Isotopes
• Glycopeptides
• Mucins
• Polysaccharides
• Sialyltransferases
• N-Acetylneuraminic Acid
• beta-Galactoside alpha-2,3-Sialyltransferase
• ST3GAL1 protein, human

Topics

• Biomarkers, Tumor (blood, chemistry, metabolism)
• Carbon Isotopes (chemistry, metabolism)
• Cell Line, Tumor
• Electrophoresis, Polyacrylamide Gel
• Female
• Glycopeptides (chemistry, metabolism)
• Humans
• Male
• Mucins (blood, chemistry, metabolism)
• N-Acetylneuraminic Acid (chemistry, metabolism)
• Neoplasms (chemistry, metabolism)
• Polysaccharides (analysis, chemistry, metabolism)
• Sialyltransferases (metabolism)
• beta-Galactoside alpha-2,3-Sialyltransferase