Mining of serum glycoproteins by an indirect approach using cell line secretome

Title
Mining of serum glycoproteins by an indirect approach using cell line secretome
Authors
안영희강운범김준이철주
Keywords
glycoprotein; LC-MS/MS; plasma proteome database; proteome; secreted protein database; secretome
Issue Date
2010-02
Publisher
Molecules and cells
Citation
VOL 29, NO 2, 123-130
Abstract
Glycosylation is the most important and abundant post-translational modification in serum proteome. Several specific types of glycan epitopes have been shown to be associated with various types of disease. Direct analysis of serum glycoproteins is challenging due to its wide dynamic range. Alternatively, glycoproteins can be disco- vered in the secretome of model cell lines and then con- firmed in blood. However, there has been little experi-men- tal evidence showing cell line secretome as a tractable target for the study of serum glycoproteins. We used a hydrazine-based glycocapture method to selectively enrich glycoproteins from the secretome of the breast cancer cell line Hs578T. A total of 132 glycoproteins were identified by nanoLC-MS/MS analysis. Among the identified proteins, we selected 13 proteins that had one or more N-gly-cosylation motifs in the matched peptides, which were included in the Secreted Protein Database but not yet in the Plasma Proteome Database (PPD), and whose antibodies were commercially available. Nine out of the 13 selected proteins were detected from human blood plasma by western analysis. Furthermore, eight proteins were also detected from the plasma by targeted LC-MS/MS, which had never been previously identified by data-dependent LC-MS/MS. Our results provide novel proteins that should be enrolled in PPD and suggest that analysis of cell line secretome with subfractionation is an efficient strategy for discovering disease-relevant serum proteins.
URI
http://pubs.kist.re.kr/handle/201004/45564
ISSN
1016-8478
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KIST Publication > Article
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