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대한화학회 제124회 학술발표회 및 총회 Extensive proteome profiling of IDH1 mutated U87MG cell line for investigating the tumorigenic roles in glioblastoma

등록일
2019년 8월 29일 16시 57분 33초
접수번호
1897
발표코드
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발표시간
10월 18일 (금요일) 11:00~12:30
발표형식
포스터
발표분야
Analytical Chemistry
저자 및
공동저자
Seunghoon Back, Jiwon Hong, Chaewon Kang1, Su-Jin Kim, Sang-Won Lee*
Department of Chemistry, Korea University, Korea
1Korea University, Korea
Glioblastoma is the lethal type of brain cancer. A recent genomewide mutational analysis of glioblastoma revealed IDH1 mutation was associated with an increase in overall survival in patients. However, proteome research related with the IDH1 mutation have not been systematically reported. Here, we present comprehensive proteomic analysis of IDH1 R132H point mutated U87MG cell line for investigating the tumorigenic roles in glioblastoma. For deep proteome profiling, 6-plex TMT labeling was performed on day 1 and day 6 samples: each of wild type, mutant 9-6 and mutant 9-19 cell line respectively. For phospho analysis, IMAC enrichment was performed on pooled 2.4 mg peptide samples and the flow-through was divided into 24 fractions using mid-pH RPLC fractionation. Phosphopeptide analysis was performed on Q Exactive HF-X which was coupled to a DO-2D-NCFC-RP/RPLC system and 24 fractionated global peptide samples were analyzed by Q Exactive HF-X which was coupled to a DO-RPLC system. An extensive profiling generated comprehensive proteome information including avg. 259,060 peptides, 11,868 protein groups and avg. 51,595 phosphopeptide. 7,352 protein groups for global and phospho analysis respectively.

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