|
Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Life Science / Faculty of Advanced Life Science >
Peer-reviewed Journal Articles, etc >
Selective reaction monitoring approach using structure-defined synthetic glycopeptides for validating glycopeptide biomarkers pre-determined by bottom-up glycoproteomics
| Title: | Selective reaction monitoring approach using structure-defined synthetic glycopeptides for validating glycopeptide biomarkers pre-determined by bottom-up glycoproteomics |
| Authors: | Shiratori, Kouta Browse this author | | Yokoi, Yasuhiro Browse this author | | Wakui, Hajime Browse this author | | Hirane, Nozomi Browse this author | | Otaki, Michiru Browse this author | | Hinou, Hiroshi Browse this author | | Yoneyama, Tohru Browse this author | | Hatakeyama, Shingo Browse this author | | Kimura, Satoshi Browse this author | | Ohyama, Chikara Browse this author | | Nishimura, Shin-Ichiro Browse this author →KAKEN DB |
| Issue Date: | 3-Aug-2022 |
| Publisher: | Royal Society of Chemistry |
| Journal Title: | RSC advances |
| Volume: | 12 |
| Issue: | 33 |
| Start Page: | 21385 |
| End Page: | 21393 |
| Publisher DOI: | 10.1039/d2ra02903k |
| Abstract: | Clusterin is a heavily glycosylated protein that is upregulated in various cancer and neurological diseases. The findings by the Hancock and Iliopoulos group that levels of the tryptic glycopeptide derived from plasma clusterin, (372)Leu-Ala-Asn-Leu-Thr-Gln-Gly-Glu-Asp-Gln-Tyr-Tyr-Leu-Arg(385) with a biantennary disialyl N-glycan (A2G2S2 or FA2G2S2) at Asn374 differed significantly prior to and after curative nephrectomy for clear cell renal cell carcinoma (RCC) patients motivated us to verify the feasibility of this glycopeptide as a novel biomarker of RCC. To determine the precise N-glycan structure attached to Asn374, whether A2G2S2 is composed of the Neu5Ac alpha 2,3Gal or/and the Neu5Ac alpha 2,6Gal moiety, we synthesized key glycopeptides having one of the two putative isomers. Selective reaction monitoring assay using synthetic glycopeptides as calibration standards allowed top-down glycopeptidomics for the absolute quantitation of targeted label-free glycopeptides in a range from 313.3 to 697.5 nM in the complex tryptic digests derived from serum samples of RCC patients and healthy controls. Our results provided evidence that the Asn374 residue of human clusterin is modified dominantly with the Neu5Ac alpha 2,6Gal structure and the levels of clusterin bearing an A2G2S2 with homo Neu5Ac alpha 2,6Gal terminals at Asn374 decrease significantly in RCC patients as compared with healthy controls. The present study elicits that a new strategy integrating the bottom-up glycoproteomics with top-down glycopeptidomics using structure-defined synthetic glycopeptides enables the confident identification and quantitation of the glycopeptide targets pre-determined by the existing methods for intact glycopeptide profiling. |
| Type: | article |
| URI: | http://hdl.handle.net/2115/86628 |
| Appears in Collections: | 生命科学院・先端生命科学研究院 (Graduate School of Life Science / Faculty of Advanced Life Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
|
|
