Selective reaction monitoring approach using structure-defined synthetic glycopeptides for validating glycopeptide biomarkers pre-determined by bottom-up glycoproteomics

Shiratori, Kouta; Yokoi, Yasuhiro; Wakui, Hajime; Hirane, Nozomi; Otaki, Michiru; Hinou, Hiroshi; Yoneyama, Tohru; Hatakeyama, Shingo; Kimura, Satoshi; Ohyama, Chikara; Nishimura, Shin-Ichiro

Selective reaction monitoring approach using structure-defined synthetic glycopeptides for validating glycopeptide biomarkers pre-determined by bottom-up glycoproteomics

Shiratori, Kouta; Yokoi, Yasuhiro; Wakui, Hajime et al.

RSC advances, 2022, 12(33), 21385-21393

Permalink : https://hdl.handle.net/2115/86628

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https://doi.org/10.1039/d2ra02903k

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アクセス権	metadata only access
URI	https://hdl.handle.net/2115/86628
タイトル	Selective reaction monitoring approach using structure-defined synthetic glycopeptides for validating glycopeptide biomarkers pre-determined by bottom-up glycoproteomics
著者	Shiratori, Kouta ; Yokoi, Yasuhiro ; Wakui, Hajime ; Hirane, Nozomi ; Otaki, Michiru ; Hinou, Hiroshi ; Yoneyama, Tohru ; Hatakeyama, Shingo ; Kimura, Satoshi ; Ohyama, Chikara ; Nishimura, Shin-Ichiro
著者名 Shiratori, Kouta
著者名 Yokoi, Yasuhiro
著者名 Wakui, Hajime
著者名 Hirane, Nozomi
著者名 Otaki, Michiru
著者名 Hinou, Hiroshi
著者名 Yoneyama, Tohru
著者名 Hatakeyama, Shingo
著者名 Kimura, Satoshi
著者名 Ohyama, Chikara
e-Rad_Researcher 00183898 著者名 Nishimura, Shin-Ichiro
言語	英語
発行日	2022-08-03
出版者	Royal Society of Chemistry
収録物名	RSC advances
巻(号)	12(33)
ページ	21385-21393
抄録	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.
資源タイプ	学術雑誌論文
出版タイプ	Version of Record
関連情報 (isIdenticalTo)	出版社版DOI https://doi.org/10.1039/d2ra02903k