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Title: Advancements and Application of Microsecond Synchrotron X-ray Footprinting at the Advanced Light Source

Abstract

The method of synchrotron X-ray protein footprinting (XF-MS) is used to determine protein conformational changes, folding, protein-protein and protein-ligand interactions, providing information which is often difficult to obtain using X-ray crystallography and other common structural biology methods [1–3]. The technique uses comparative in situ labeling of solvent-accessible side chains by highly reactive hydroxyl radicals (•OH) in buffered aqueous solution under different assay conditions. In regions where a protein is folded or binds a partner, these •OH susceptible sites are inaccessible to solvent, and therefore protected from labeling. The •OH are generated by the ionization of water using high-flux-density X-rays. High-flux density is a key factor for XF-MS labeling because obtaining an adequate steady-state concentration of hydroxyl radical within a short irradiation time is necessary to minimize radiation-induced secondary damage and also to overcome various scavenging reactions that reduce the yield of labeled side chains.

Authors:
; ; ;
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1576495
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Synchrotron Radiation News
Additional Journal Information:
Journal Volume: 29; Journal Issue: 1; Journal ID: ISSN 0894-0886
Publisher:
Taylor & Francis
Country of Publication:
United States
Language:
English

Citation Formats

Gupta, Sayan, Celestre, Rich, Feng, Jun, and Ralston, Corie. Advancements and Application of Microsecond Synchrotron X-ray Footprinting at the Advanced Light Source. United States: N. p., 2016. Web. doi:10.1080/08940886.2016.1124684.
Gupta, Sayan, Celestre, Rich, Feng, Jun, & Ralston, Corie. Advancements and Application of Microsecond Synchrotron X-ray Footprinting at the Advanced Light Source. United States. doi:10.1080/08940886.2016.1124684.
Gupta, Sayan, Celestre, Rich, Feng, Jun, and Ralston, Corie. Sat . "Advancements and Application of Microsecond Synchrotron X-ray Footprinting at the Advanced Light Source". United States. doi:10.1080/08940886.2016.1124684. https://www.osti.gov/servlets/purl/1576495.
@article{osti_1576495,
title = {Advancements and Application of Microsecond Synchrotron X-ray Footprinting at the Advanced Light Source},
author = {Gupta, Sayan and Celestre, Rich and Feng, Jun and Ralston, Corie},
abstractNote = {The method of synchrotron X-ray protein footprinting (XF-MS) is used to determine protein conformational changes, folding, protein-protein and protein-ligand interactions, providing information which is often difficult to obtain using X-ray crystallography and other common structural biology methods [1–3]. The technique uses comparative in situ labeling of solvent-accessible side chains by highly reactive hydroxyl radicals (•OH) in buffered aqueous solution under different assay conditions. In regions where a protein is folded or binds a partner, these •OH susceptible sites are inaccessible to solvent, and therefore protected from labeling. The •OH are generated by the ionization of water using high-flux-density X-rays. High-flux density is a key factor for XF-MS labeling because obtaining an adequate steady-state concentration of hydroxyl radical within a short irradiation time is necessary to minimize radiation-induced secondary damage and also to overcome various scavenging reactions that reduce the yield of labeled side chains.},
doi = {10.1080/08940886.2016.1124684},
journal = {Synchrotron Radiation News},
number = 1,
volume = 29,
place = {United States},
year = {2016},
month = {1}
}

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