Summary
Changes in N-linked glycosylation are known to occur during the development of cancer. For example, increased branching of oligosaccharides has been associated with metastasis and has been correlated to tumor progression in human cancers of the breast, colon, and melanomas. Increases in core fucosylation have also been associated with the development of hepatocellular carcinoma (HCC). To a large extent, the proteins to which these N-linked glycans are attached have been unknown. However, with the advent of sensitive glycan analysis and proteomic technologies, the ability to comprehensively identify all the fucosylated proteins in a given population is now a possibility. This method, generally referred to as targeted glycoproteomics, is shown as applied to the detection of proteins present in the fucosylated proteome of a liver cancer cell line but is generally enough to be applied in many other situations.
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References
Hakomori S (1996) Tumor malignancy defined by aberrant glycosylation and sphingo(glyco)lipid metabolism. Cancer Res. 56(23): 5309–18
Kobata A (1998) A retrospective and prospective view of glycopathology. Glycoconj J. 15(4): 323–31
Dennis JW, Laferte S, Waghorne C, Breitman ML, Kerbel RS (1987) Beta 1–6 branching of Asn-linked oligosaccharides is directly associated with metastasis. Science. 236(4801): 582–5
Fernandes B, Sagman U, Auger M, Demetrio M, Denni JW (1991) Beta 1-6 branched oligosaccharides as a marker of tumor progression in human breast and colon neoplasia [see comments]. Cancer Res. 51(2): 718–23
Seelentag WK, Li WP, Schmitz SF, Metzger U, Aeberhard P, Heitz PU, Roth J (1998) Prognostic value of beta1, 6-branched oligosaccharides in human colorectal carcinoma. Cancer Res. 58(23): 5559–64
Yoshimura, M, Nishikawa A, Ihara Y, Taniguchi S, Taniguchi N (1995) Suppression of lung metastasis of B16 mouse melanoma by N-acetylglucosaminyltransferase III gene transfection. Proc Natl Acad Sci U S A. 92(19): 8754–8
Breborowicz J, Mackiewicz A, Breborowicz D (1981) Microheterogeneity of alpha-fetoprotein in patient serum as demonstrated by lectin affino-electrophoresis. Scand J Immunol. 14(1): 15–20
Miyazaki J, Endo Y, Oda T (1981) Lectin affinities of alpha-fetoprotein in liver cirrhosis, hepatocellular carcinoma and metastatic liver tumor. Acta Hepatol. Jpn. 22: 1559–68
Taketa K, Ichikawa E, Taga H, Hirai H, (1985) Antibody-affinity blotting, a sensitive technique for the detection of alpha-fetoprotein separated by lectin affinity electrophoresis in agarose gels. Electrophoresis. 6: 492–97
Taketa K, Sekiya C, Namiki M, Akamatsu K, Ohta Y, Endo Y, Kosaka K (1990) Lectin-reactive profiles of alpha-fetoprotein characterizing hepatocellular carcinoma and related conditions. Gastroenterology. 99(2): 508–18
Shiraki K, Takase K, Tameda Y, Hamada M, Kosaka Y, Nakano T (1995) A clinical study of lectin-reactive alpha-fetoprotein as an early indicator of hepatocellular carcinoma in the follow-up of cirrhotic patients. Hepatology. 22(3): 802–7
Hutchinson WL, Du MQ, Johnson PJ, Williams R (1991) Fucosyltransferases: differential plasma and tissue alterations in hepatocellular carcinoma and cirrhosis. Hepatology. 13(4): 683–8
Miyoshi E, Noda K, Yamaguchi Y, Inoue S, Ikeda Y, Wang W, Ko JH, Uozumi N, Li W, Taniguchi N (1999) Altered glycosylation of serum transferrin of patients with hepatocellular carcinoma. J Biol Chem. 264(5): 2415–23
Yamashita K, Koide N, Endo T, Iwaki Y, Kobata A (1989) Altered glycosylation of serum transferrin of patients with hepatocellular carcinoma. J Biol Chem. 264(5): 2415–23
Naitoh A, Aoyagi Y, Asakura H (1999) Highly enhanced fucosylation of serum glycoproteins in patients with hepatocellular carcinoma. J Gastroenterol Hepatol. 5(1): 43–68
Guile, G. R., P.M. Rudd, D.R. Wing, S.B. Prime, and R.A. Dwek (1996) A rapid high-resolution high-performance liquid chromatographic method for separating glycan mixtures and analyzing oligosaccharide profiles. Anal Biochem. 240(2): 21017.
Rudd PM, Mattu TS, Zitzmann N, Mehta A, Colominas C, Hart E, Opdenakker G, DwekRA (1999) Glycoproteins: rapid sequencing technology for N-linked and GPI anchor glycans. Biotechnol Genet Eng Rev. 16: 1–21
Rudd PM, Mattu TS, Masure S, Bratt T, Van den Steen PE, Wormald MR, Kuster B, Harvey DJ, Borregaard N, Van Damme J, Dwek RA, Opdenakker G (1999) Glycosylation of natural human neutrophil gelatinase B and neutrophil gelatinase B-associated lipocalin. Biochemistry. 38(42): 13937–50
Mattu TS, Royle L, Langridge J, Wormald MR, Van den Steen PE, Van Damme J, Opdenakker G, Harvey DJ, Dwek RA, Rudd, PM(2000) “O-glycan analysis of natural human neutrophil gelatinase B using a combination of normal phase-HPLC and online tandem mass spectrometry: implications for the domain organization of the enzyme. Biochemistry. 39(51): 15695–704
Rudd PM, Colominas C, Royle L, Murphy N, Hart E, Merry AH, Hebestreit HF, Dwek RA (2001) A high-performance liquid chromatography based strategy for rapid, sensitive sequencing of N-linked oligosaccharide modifications to proteins in sodium dodecyl sulphate polyacrylamide electrophoresis gel bands. Proteomics. 1(2): 285–289
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Comunale, M., Mehta, A. (2009). Glycoproteomic Analysis by Two-Dimensional Electrophoresis. In: Tainsky, M. (eds) Tumor Biomarker Discovery. Methods in Molecular Biology, vol 520. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-811-9_5
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DOI: https://doi.org/10.1007/978-1-60327-811-9_5
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