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N-Linked Glycosylation of the Liver Cancer Biomarker GP73
Journal article   Open access   Peer reviewed

N-Linked Glycosylation of the Liver Cancer Biomarker GP73

Pamela A Norton, Mary Ann Comunale, Jonathan Krakover, Lucy Rodemich, Natalie Pirog, Anthony D'Amelio, Ramila Philip, Anand S Mehta and Timothy M Block
Journal of cellular biochemistry, v 104(1)
01 May 2008
PMID: 18004786
url
https://doi.org/10.1002/jcb.21610View
Published, Version of Record (VoR) Open

Abstract

Golgi protein 73 fucosylation hepatocellular carcinoma
The association between elevated circulating levels of GP73 (and fucosylated GP73 in particular) and hepatocellular carcinoma suggests that a thorough analysis of the extent of GP73 glycosylation is warranted. Detailed analysis of the glycosylation patterns of such low abundance proteins are hampered by technical difficulties. Using conventional lectin affinity chromatography, we have established that three quarters of the GP73 secreted from a cell line derived from HCC is fucosylated. Using mass spectrometry, we have established that at least two of three potential sites of N-linked glycosylation are occupied on most molecules of GP73 secreted from cultured hepatoma cells. Furthermore, the oligosaccharides added to recombinant GP73 resemble those present in the bulk of secreted protein, mostly bi-antennary with core fucose, with a smaller fraction of tri- and tetra-antennary structures. The frequency of fucosylation observed on the recombinant protein agrees well with the pattern of lectin binding of the endogenous secreted protein. Finally, we have developed a method to interrogate the glycans added to either the near full length protein or at a particular sequon, providing proof of concept that a small peptide embedded in a heterologous context can preserve both fucosylation and a high level of branching of oligosaccharides added.

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Web of Science research areas
Biochemistry & Molecular Biology
Cell Biology
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