인문학
사회과학
자연과학
공학
의약학
농수해양학
예술체육학
복합학
DBpia AI
AI 에이전트
AI 아이디어
AI 리더
AI 뷰어
Citeasy
크롬 서지관리 다운로드
지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
초록·키워드
Telomerase reverse transcriptase is crucial for cellular development, regeneration, and disease processes. Strategies for both telomerase activation and inhibition have been intensively explored in the past decades. In this study, we present a highly miniaturized, microarray-based assay designed to identify compounds that target telomerase. The active protein was either recombinantly derived from E. coli or obtained from cell lysates of human cancer cell lines and mouse cells expressing telomerase. Using non-contact spotter technology, these lysates or purified telomerase proteins were transferred onto nitrocellulose pads on a microarray. A telomerase binding assay, incorporating fluorescent labelled primer, the template RNA telomerase RNA component, and fluorescent labelled nucleotide as a primer cocktail, was conducted in incubation chambers. Binding of this primer cocktail to spotted telomerase from cell lysates, and from purified recombinant telomerase resulted in an increase in bound fluorescence. Epigallocatechin gallate, a known telomerase inhibitor, reduced this fluorescence in a dose-dependent manner with micromolar affinity. The inhibitory effect on telomerase was validated by thermophoresis and its impact on activity was shown in a Telomerase Repeated Amplification Protocol (TRAP) assay. Additional screening identified that 4'-iodo cytochalasin H inhibits primer cocktail binding to cell lysate in the low micromolar range. Molecular modeling and docking pinpointed a putative binding site for epigallocatechin gallate in a human telomerase homologue, and a putative binding site for 4'-iodo cytochalasin H. In summary, we developed an assay that can be employed to discover new telomerase inhibitors and that will serve as a valuable tool for screening of activators.
인공지능 문자 인식 모델을 통해 추출된 텍스트로, 일부 오타나 오류가 포함될 수 있으나 지속적으로 개선 중입니다.
오류를 발견하셨다면 해당 부분을 드래그한 후 ' 를 통해 신고해주세요.
오류를 발견하셨다면 해당 부분을 드래그한 후 ' 를 통해 신고해주세요.
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