기술이전

 

탄뎀질량분석기를 이용한 인체내 O-형 당펩티드의 분석법

관리번호
RS05202012210001
첨부파일
개발년도
2020
저자정보
유종신
참고문헌
Anal. Chem. 2020, 92, 9772−9781
관련장비
하이브리드 FT-ETD 질량분석기
응용분야
인간 혈액및 소변내 O-형 당단백질 분석
연구분야
BT
분석/적용분야
BT
기술분야
질량분석기술
분석/공동연구지원
인간 혈액내 암관련 당단백질체 발굴 인간 소변내 질병관련 O-형 당단백질 발굴
요약
We present an O-GlycoProteome Analyzer (O-GPA) that automatically classifies mucin-type O-glycosylation using higherenergy collisional dissociation (HCD) in mass spectrometry.
키워드
탄뎀질량분석, O-형 당펩티드, 당단백질 O?Glycopeptides, Higher-EnergyCollisional Dissociation, Mass Spectrometry
연구개발성과
질량분석기를 이용한 당단백질 분석법
내 용
Changes in mucin-type O-glycosylation of human proteins affect protein function, immune response, and cancer progression. Since O-glycoproteins are characterized by the microheterogeneity of diverse O-glycans with no conserved sequence and the macroheterogeneity of multiple glycosylation sites on serine and/or threonine in human proteins, the assessment of different mucin types, such as Tn-antigen, core 1, and core 2, and their extended core types in O-glycopeptides, is extremely challenging.
Here, we present an O-GlycoProteome Analyzer (O-GPA) that automatically classifies mucin-type O-glycosylation using higherenergy collisional dissociation (HCD) in mass spectrometry. First, we estimated the number of GlcNAc residues using the intensity ratio of GlcNAc-specific fragment ions (HexNAc-CH6O3 and HexNAc-2H2O) over GalNAc-specific fragment ions (HexNAcC2H6O3 and HexNAc-C2H4O2) in the HCD spectrum. Furthermore, we classified the different mucin types of O-glycopeptides from characteristic B2 (HexNAc2) or Y2α (PEP + HexNAc2), and Y2β (PEP + HexNAcHex) fragment ions, along with the number of GlcNAc. Furthermore, O-GPA automatically determined single or multiple O-glycosylation, regardless of the mucin types. The mucin type of O-glycopeptides from human urine and plasma was confirmed with an overall accuracy of 96%. We found 97 core 1, 56 core 2, 13 extended core 1, and 12 extended core 2 glycopeptides from urine; and 22 core 1, 13 core 2, 7 extended core 1, 1 extended core 2, and 1 Tn-antigen from plasma. Our strategy can be used to successfully characterize specific mucin types of Oglycoproteins in human biological samples.