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제116회 대한화학회 학술발표회, 총회 및 기기전시회 안내 Rational design of a photo-crosslinking BODIPY for in situ protein labeling

등록일
2015년 9월 3일 14시 45분 33초
접수번호
1246
발표코드
ORGN.P-422 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
발표시간
10월 15일 (목요일) 11:00~12:30
발표형식
포스터
발표분야
유기화학
저자 및
공동저자
Murale Dhiraj, 홍성철1, 이준석2,*
한국과학기술연구원(KIST) 분자인식연구센터, Korea
1과학기술연합대학원대학교(UST) 생물화학, Korea
2한국과학기술연구원(KIST) 분자인지연구센터, Korea

Photo-crosslinking agents have emerged as critical tools to investigate protein-protein interactions in complex proteome, but there are few photocrosslinkers available at the moment. Here, we report the first rational design of a photo-crosslinking BODIPY fluorophore (pcBD) and its biological application for biomolecule labeling in a spatiotemporally controlled manner. As a photosensitizing functional motif, the aryl ketone group was incorporated into BODIPY fluorophore, and series of proteins were labeled by pcBD compounds upon UV irradiations. Particularly, these probes could crosslink proteins under both 254 nm and 365 nm UV lights, though they generally exhibited high crosslinking efficiency under 254 nm light conditions. The chief advantage of this BODIPY-benzophenone conjugate system for photo-activated protein labeling is the capability to attach BODIPY fluorophore at any kinds of target proteins depending on the spatial vicinity. Here we chose ubiquitin ligase complex as a model system since several enzymes tightly cooperate to tag substrate proteins with ubiquitin during the ubiquitination process. We have synthesized a photo-affinity probe, pcBD-E3binder, by conjugating E3 ligase binding peptide and BODIPY-benzophenone conjugate. As a result of this experiment we proved the utility of photo-crosslinking fluorophore for spatiotemporal protein labeling in complex mixtures. In our future work we want to use this BODIPY-benzophenone conjugate probe to tag and identify the substrates for ubiquitin ligases along with their binding partners.


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