Mol. Cells 2010; 29(2): 123-130
Published online December 10, 2009
https://doi.org/10.1007/s10059-010-0008-0
© The Korean Society for Molecular and Cellular Biology
Correspondence to : *Correspondence: clee270@kist.re.kr
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.
Keywords , LC-MS/MS, plasma proteome database, proteome, secreted protein database, secretome
Mol. Cells 2010; 29(2): 123-130
Published online February 28, 2010 https://doi.org/10.1007/s10059-010-0008-0
Copyright © The Korean Society for Molecular and Cellular Biology.
Younghee Ahn1,3, Un-Beom Kang1,2,3, Joon Kim2, and Cheolju Lee1,*
1Life Sciences Division, Korea Institute of Science and Technology, Seoul 136-791, Korea, 2School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Korea, 3These authors contributed equally to this work.
Correspondence to:*Correspondence: clee270@kist.re.kr
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.
Keywords: , LC-MS/MS, plasma proteome database, proteome, secreted protein database, secretome
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