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Title: Development of Container Free Sample Exposure for Synchrotron X-ray Footprinting

Abstract

The method of X-ray footprinting and mass spectrometry (XFMS) on large protein assemblies and membrane protein samples requires high flux density to overcome the hydroxyl radical scavenging reactions produced by the buffer constituents and the total protein content. Previously, we successfully developed microsecond XFMS using microfluidic capillary flow and a microfocused broadband X-ray source at the Advanced Light Source synchrotron beamlines, but the excessive radiation damage incurred when using capillaries prevented the full usage of a high-flux density beam. Here we present another significant advance for the XFMS method: the instrumentation of a liquid injection jet to deliver container free samples to the X-ray beam. Our preliminary experiments with a liquid jet at a bending magnet X-ray beamline demonstrate the feasibility of the approach and show a significant improvement in the effective dose for both the Alexa fluorescence assay and protein samples compared to conventional capillary flow methods. The combination of precisely controlled high dose delivery, shorter exposure times, and elimination of radiation damage due to capillary effects significantly increases the signal quality of the hydroxyl radical modification products and the dose–response data. This new approach is the first application of container free sample handling for XFMS and opens upmore » the method for even further advances, such as high-quality microsecond time-resolved XFMS studies.« less

Authors:
 [1];  [1]; ORCiD logo [1];  [2]; ORCiD logo [1]
  1. Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
  2. Stanford Linear Accelerator Center, Menlo Park, California 94025, United States
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
OSTI Identifier:
1591700
Alternate Identifier(s):
OSTI ID: 1605365; OSTI ID: 1756324
Grant/Contract Number:  
AC02-05CH11231; 1R01GM126218; AC02-76SF00515
Resource Type:
Published Article
Journal Name:
Analytical Chemistry
Additional Journal Information:
Journal Name: Analytical Chemistry Journal Volume: 92 Journal Issue: 1; Journal ID: ISSN 0003-2700
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Hydroxyls; x-rays; modification; liquids; peptides and proteins

Citation Formats

Gupta, Sayan, Chen, Yan, Petzold, Christopher J., DePonte, Daniel P., and Ralston, Corie Y. Development of Container Free Sample Exposure for Synchrotron X-ray Footprinting. United States: N. p., 2019. Web. doi:10.1021/acs.analchem.9b04849.
Gupta, Sayan, Chen, Yan, Petzold, Christopher J., DePonte, Daniel P., & Ralston, Corie Y. Development of Container Free Sample Exposure for Synchrotron X-ray Footprinting. United States. https://doi.org/10.1021/acs.analchem.9b04849
Gupta, Sayan, Chen, Yan, Petzold, Christopher J., DePonte, Daniel P., and Ralston, Corie Y. Mon . "Development of Container Free Sample Exposure for Synchrotron X-ray Footprinting". United States. https://doi.org/10.1021/acs.analchem.9b04849.
@article{osti_1591700,
title = {Development of Container Free Sample Exposure for Synchrotron X-ray Footprinting},
author = {Gupta, Sayan and Chen, Yan and Petzold, Christopher J. and DePonte, Daniel P. and Ralston, Corie Y.},
abstractNote = {The method of X-ray footprinting and mass spectrometry (XFMS) on large protein assemblies and membrane protein samples requires high flux density to overcome the hydroxyl radical scavenging reactions produced by the buffer constituents and the total protein content. Previously, we successfully developed microsecond XFMS using microfluidic capillary flow and a microfocused broadband X-ray source at the Advanced Light Source synchrotron beamlines, but the excessive radiation damage incurred when using capillaries prevented the full usage of a high-flux density beam. Here we present another significant advance for the XFMS method: the instrumentation of a liquid injection jet to deliver container free samples to the X-ray beam. Our preliminary experiments with a liquid jet at a bending magnet X-ray beamline demonstrate the feasibility of the approach and show a significant improvement in the effective dose for both the Alexa fluorescence assay and protein samples compared to conventional capillary flow methods. The combination of precisely controlled high dose delivery, shorter exposure times, and elimination of radiation damage due to capillary effects significantly increases the signal quality of the hydroxyl radical modification products and the dose–response data. This new approach is the first application of container free sample handling for XFMS and opens up the method for even further advances, such as high-quality microsecond time-resolved XFMS studies.},
doi = {10.1021/acs.analchem.9b04849},
journal = {Analytical Chemistry},
number = 1,
volume = 92,
place = {United States},
year = {2019},
month = {12}
}

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