An automated liquid jet for fluorescence dosimetry and microsecond radiolytic labeling of proteins
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Published:2022-08-25
Issue:1
Volume:5
Page:
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ISSN:2399-3642
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Container-title:Communications Biology
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language:en
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Short-container-title:Commun Biol
Author:
Rosi Matthew, Russell Brandon, Kristensen Line G., Farquhar Erik R.ORCID, Jain RohitORCID, Abel Donald, Sullivan Michael, Costello Shawn M.ORCID, Dominguez-Martin Maria Agustina, Chen Yan, Marqusee SusanORCID, Petzold Christopher J.ORCID, Kerfeld Cheryl A.ORCID, DePonte Daniel P., Farahmand Farid, Gupta SayanORCID, Ralston Corie Y.ORCID
Abstract
AbstractX-ray radiolytic labeling uses broadband X-rays for in situ hydroxyl radical labeling to map protein interactions and conformation. High flux density beams are essential to overcome radical scavengers. However, conventional sample delivery environments, such as capillary flow, limit the use of a fully unattenuated focused broadband beam. An alternative is to use a liquid jet, and we have previously demonstrated that use of this form of sample delivery can increase labeling by tenfold at an unfocused X-ray source. Here we report the first use of a liquid jet for automated inline quantitative fluorescence dosage characterization and sample exposure at a high flux density microfocused synchrotron beamline. Our approach enables exposure times in single-digit microseconds while retaining a high level of side-chain labeling. This development significantly boosts the method’s overall effectiveness and efficiency, generates high-quality data, and opens up the arena for high throughput and ultrafast time-resolved in situ hydroxyl radical labeling.
Funder
U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences
Publisher
Springer Science and Business Media LLC
Subject
General Agricultural and Biological Sciences,General Biochemistry, Genetics and Molecular Biology,Medicine (miscellaneous)
Reference52 articles.
1. Gupta, S., Celestre, R., Petzold, C. J., Chance, M. R. & Ralston, C. Development of a microsecond X-ray protein footprinting facility at the advanced light source. J. Synchrotron Radiat. 21, 690–699 (2014). 2. Xu, G. & Chance, M. R. Hydroxyl radical-mediated modification of proteins as probes for structural proteomics. Chem. Rev. 107, 3514–3543 (2007). 3. Gupta, S., Feng, J., Chan, L. J., Petzold, C. J. & Ralston, C. Y. Synchrotron X-ray footprinting as a method to visualize water in proteins. J. Synchrotron Radiat. 23, 1056–1069 (2016). 4. Kiselar, J. G., Maleknia, S. D., Sullivan, M., Downard, K. M. & Chance, M. R. Hydroxyl radical probe of protein surfaces using synchrotron X-ray radiolysis and mass spectrometry. Int J. Radiat. Biol. 78, 101–114 (2002). 5. Shcherbakova, I., Mitra, S., Beer, R. H. & Brenowitz, M. Fast Fenton footprinting: a laboratory-based method for the time-resolved analysis of DNA, RNA and proteins. Nucleic Acids Res. 34, e48 (2006).
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