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식물노화·수명연구단
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Photo-Activated Crosslinking Mass Spectrometry for Studying Biomolecular Interactions

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Title
Photo-Activated Crosslinking Mass Spectrometry for Studying Biomolecular Interactions
Author(s)
Samuel Kim; Jae Kyoo Lee; Hong Gil Nam; Richard N. Zare
Publication Date
2014-01
Journal
BIOPHYSICAL JOURNAL, v.106, no.2, pp.459A - 459A
Publisher
CELL PRESS
Abstract
The structural and mechanistic analysis of biomolecular interaction is important for understanding the molecular basis of a wide range of biological phenomenon. Relatively weak interactions between functional molecules may play crucial roles in regulating highly networked and dynamically controlled biological systems. This type of interaction, however, is more challenging to study because of its transient nature. As a general solution to the problem, we propose the use of chemical crosslinking in combination with mass spectrometry and apply it to protein-protein interactions. Ultraviolet-activatable crosslinker molecules that are incorporated to the protein of interest can be utilized to capture transient interactions under a physiological condition by forming covalent bonds between interacting molecules in close proximity via carbene chemistry. The crosslinked sample is then enzymatically digested and analyzed by liquid chromatography-mass spectrometry. High mass resolution analysis ensures identification of crosslinked peptide species and enables spatial mapping onto available structural models obtained from crystallography or NMR spectroscopy. The proposed methodology is demonstrated with a model system of cytochrome c and its oxidase, where we find multiple binding modes and explore their possible role in controlling enzymatic activity.
URI
https://pr.ibs.re.kr/handle/8788114/798
DOI
10.1016/j.bpj.2013.11.2601
ISSN
0006-3495
Appears in Collections:
Center for Plant Aging Research (식물 노화·수명 연구단) > 1. Journal Papers (저널논문)
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