Earticle

초록

영어

Glycoprotein conformations are complex and heterogeneous. Currently, site-specific characterization of glycopeptides is a challenge. We sought to establish an efficient method of N-glycoprotein characterization using mass spectrometry (MS). Using alpha-1-acid glycoprotein (AGP) as a model N-glycoprotein, we identified its tryptic N-glycopeptides and examined the data reproducibility in seven laboratories running different LC−MS/MS platforms. We used three test samples and one blind sample to evaluate instrument performance with entire sample preparation workflow. 165 site-specific N-glycopeptides representative of all N-glycosylation sites were identified from AGP 1 and AGP 2 isoforms. The glycopeptide fragmentations by collision-induced dissociation or higher-energy collisional dissociation (HCD) varied based on the MS analyzer. Orbitrap Elite identified the greatest number of AGP N-glycopeptides, followed by Triple TOF and Q-Exactive Plus. Reproducible generation of oxonium ions, glycancleaved glycopeptide fragment ions, and peptide backbone fragment ions was essential for successful identification. Laboratory proficiency affected the number of identified N-glycopeptides. The relative quantities of the 10 major N-glycopeptide isoforms of AGP detected in four laboratories were compared to assess reproducibility. Quantitative analysis showed that the coefficient of variation was <25% for all test samples. Our analytical protocol yielded identification and quantification of site-specific N-glycopeptide isoforms of AGP from control and disease plasma sample.

저자

• Ju Yeon Lee [ Biomedical Omics Group, Korea Basic Science Institute, Ochang, Republic of Korea ]
• Hyun Kyoung Lee [ Biomedical Omics Group, Korea Basic Science Institute, Ochang, Republic of Korea, Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, Republic of Korea ]
• Gun Wook Park [ Biomedical Omics Group, Korea Basic Science Institute, Ochang, Republic of Korea, Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, Republic of Korea ]
• Heeyoun Hwang [ Biomedical Omics Group, Korea Basic Science Institute, Ochang, Republic of Korea ]
• Hoi Keun Jeong [ Biomedical Omics Group, Korea Basic Science Institute, Ochang, Republic of Korea, Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, Republic of Korea ]
• Ki Na Yun [ Biomedical Omics Group, Korea Basic Science Institute, Ochang, Republic of Korea, Department of Chemistry, Sogang University, Seoul, Republic of Korea ]
• Eun SunJ i [ Biomedical Omics Group, Korea Basic Science Institute, Ochang, Republic of Korea, Department of Chemistry, Hannam University, Daejeon, Republic of Korea ]
• Kwang Hoe Kim [ Biomedical Omics Group, Korea Basic Science Institute, Ochang, Republic of Korea, Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, Republic of Korea ]
• Jun Seok Kim [ Department of Biomedical Systems Engineering, Korea Polytechnics ]
• Jong Won Kim [ New Drug Development Center, Osong Medical Innovation Foundation ]
• Sung Ho Yun [ Drug & Disease Target Group, Korea Basic Science Institute, Daejeon, Republic of Korea ]
• Chi-Won Choi [ Drug & Disease Target Group, Korea Basic Science Institute, Daejeon, Republic of Korea ]
• Seung Il Kim [ Drug & Disease Target Group, Korea Basic Science Institute, Daejeon, Republic of Korea ]
• Jong-Sun Lim [ Yonsei Proteome Research Center, Yonsei University, Seoul, Republic of Korea ]
• Seul-Ki Jeong [ Yonsei Proteome Research Center, Yonsei University, Seoul, Republic of Korea ]
• Young-Ki Paik [ Yonsei Proteome Research Center, Yonsei University, Seoul, Republic of Korea ]
• Soo-Youn Lee [ Department of Laboratory & Genetics, Samsung Medical Center, Sungkyunkwan University of Medicine, Seoul, Korea, Department of Clinical Pharmacology and Therapeutics, Samsung Medical Center, Seoul, South Korea ]
• Jisook Park [ Samsung Biomedical Research Institute, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea ]
• Su Yeon Kim [ Department of Clinical Research Supporting Team, Clinical Research Institute, Samsung Medical Center, Seoul, Korea ]
• Young-Jin Choi [ Department of Biochemistry, BK21 PLUS Program for Creative Veterinary Science Research and Research Institute ]
• Yong-In Kim [ Department of Biochemistry, BK21 PLUS Program for Creative Veterinary Science Research and Research Institute ]
• Jawon Seo [ Department of Biochemistry, BK21 PLUS Program for Creative Veterinary Science Research and Research Institute ]
• Je-Yoel Cho [ Department of Biochemistry, BK21 PLUS Program for Creative Veterinary Science Research and Research Institute ]
• Myoung Jin Oh [ Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, Republic of Korea ]
• Nari Seo [ Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, Republic of Korea ]
• Hyun Joo An [ Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, Republic of Korea ]
• Jin Young Kim [ Biomedical Omics Group, Korea Basic Science Institute, Ochang, Republic of Korea ]
• Jong Shin Yoo [ Biomedical Omics Group, Korea Basic Science Institute, Ochang, Republic of Korea, Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, Republic of Korea ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국당과학회 학술대회

• 간기

  연간

• 수록기간

  2006~2022

• 십진분류

  KDC 517 DDC 614