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Three-color single molecule fluorescence resonance energy transfer: a new method for structures and dynamics of biomolecules

등록일
2005년 2월 17일 15시 44분 07초
접수번호
0988
발표코드
21P99포 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
발표시간
토 <발표Ⅲ>
발표형식
포스터
발표분야
물리화학
저자 및
공동저자
이남기, 고혜란, 김성근
서울대학교 화학부,
Analysis of structure, dynamics, and interactions of biomolecules is crucial for understanding biological mechanisms. Single-molecule fluorescence resonance energy transfer (sm-FRET) has become a powerful tool for the conformational and dynamical studies of biomolecules. These studies have been so far limited to a “two-color system” for a donor-acceptor pair of fluorophores, however, where only one distance is observed. Since most biological phenomena involve reactions and conformational changes of more than one building block of a macromolecular system, observation of multiple distances is in great demand. We developed a general method to measure three distances at a time by observing 3-color FRET at the single-molecule level, which was achieved by extending the recently developed alternating-laser-excitation (ALEX) technique to a 3-color level. ALEX is a novel technique to perform fluorescence-aided molecular sorting of fluorescent species according to their energy transfer efficiency as well as their labeling states. Using 3-color ALEX, we were able to sort triply-labeled species out of a heterogeneous mixture, and quantitatively measured three distances at a time.

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