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:
-
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Advanced Light Source (ALS)
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- 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
- Subject:
- 47 OTHER INSTRUMENTATION
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. Mon .
"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 = {2}
}
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