Dasom An, Hyejin Kim, Jin Young Kim, Jong Shin Yoo, Heeyoun Hwang
언어
영어(ENG)
URL
https://www.earticle.net/Article/A415485
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원문정보
초록
영어
O-GlcNAcylation is one of the post-translational modification that occurs at Ser/Thr residues of a protein by O-GlcNAc transferase (OGT), but the O-GlcNAcylation is regulated by O-GlcNAcase (OGA). The O-GlcNAcylation is involved in various cellular processes and diseases including diabetes, cancer and Alzheimer’s disease. Herein, we suggest a new method for identifying O-GlcNAcylated proteins including classification of O-GlcNAc and O-GalNAc with the same molecular weight. Using the fractionated raw files of LC-MS/MS from T24 cell lines, we developed an identification algorithm coded by python 3.9 as follows: 1.Selection of spectra with oxonium ions in ms2 file extracted from raw MS data obtained using LC-MS/MS; 2.Spectral matching with 20 ppm against a tryptic and O-GlcNAcylated peptide database generated from proteins identified using IP2 proteomic search result; 3.In the candidates, matching between tandem MS with b/y and c/z ion list and experimental HCD, ETD spectra, respectively; 4.Selection of the best scored O-GlcNAcylated peptide from the candidated peptides calculated by scoring system; 5.Classification of O-GlcNAc and O-GalNAc based on their corresponding fragment ions; 6.Evaluating a model using ROC curve comparing search results with manual validation. In addition, we also performed relative quantitative analysis in conjunction with the TMT Quantification method. In the glycoscience field, our study could be applied to various human disease treatments by identifying O-GlcNAcylated proteins and by engaging in cell signaling after regulating the function of them.
저자
Dasom An [ Research Center for Bioconvergence Analysis, Korea Basic Science Institute ]
Hyejin Kim [ Research Center for Bioconvergence Analysis, Korea Basic Science Institute, Department of Bio and Brain Engineering, KAIST, Daejeon 34141, Republic of Korea ]
Jin Young Kim [ Research Center for Bioconvergence Analysis, Korea Basic Science Institute ]
Jong Shin Yoo [ Research Center for Bioconvergence Analysis, Korea Basic Science Institute, Department of Bio and Brain Engineering, KAIST, Daejeon 34141, Republic of Korea ]
Heeyoun Hwang [ Research Center for Bioconvergence Analysis, Korea Basic Science Institute ]
Corresponding Author
본 학회는 화학, 생화학, 분자생물학, 미생물학, 식품공학, 의학, 약학, 유전공학 및 생물공학, 환경 및 기타 공업 등 전 분야의 탄수화물관련 이론과 기술을 연구 발전시키고 산학협동을 통해 이를 보급하여 국내 관련 산업의 발전 및 국민생활의 과학화에 기여하고자 하며, 이러한 목표와 비젼의 실현을 위해 회원들이 적극적인 참여와 활동을 전개하고자 한다.