Ahn, Younghee Kang, Un-Beom Kim, Joon Lee, Cheolju 2024-01-20T20:01:47Z 2024-01-20T20:01:47Z 2021-09-02 2010-02 1016-8478 https://pubs.kist.re.kr/handle/201004/131767 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 discovered in the secretome of model cell lines and then confirmed in blood. However, there has been little experi-mental 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-glycosylation 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. English KOREAN SOC MOLECULAR & CELLULAR BIOLOGY BREAST-CANCER QUANTITATIVE PROTEOMICS MASS-SPECTROMETRY LIQUID-CHROMATOGRAPHY HYDRAZIDE CHEMISTRY N-GLYCOSYLATION PLASMA PROTEOME IDENTIFICATION PROTEINS BIOMARKER Mining of serum glycoproteins by an indirect approach using cell line secretome Article 10.1007/s10059-010-0008-0 1 MOLECULES AND CELLS, v.29, no.2, pp.123 - 130 MOLECULES AND CELLS 29 2 123 130 scie scopus kci ART001420933 000275123100004 2-s2.0-77949424965 Biochemistry & Molecular Biology Cell Biology Biochemistry & Molecular Biology Cell Biology Article BREAST-CANCER QUANTITATIVE PROTEOMICS MASS-SPECTROMETRY LIQUID-CHROMATOGRAPHY HYDRAZIDE CHEMISTRY N-GLYCOSYLATION PLASMA PROTEOME IDENTIFICATION PROTEINS BIOMARKER glycoprotein LC-MS/MS plasma proteome database proteome secreted protein database secretome