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Title: A Mo-anode-based in-house source for small-angle X-ray scattering measurements of biological macromolecules

Abstract

We demonstrate the use of a molybdenum-anode-based in-house small-angle X-ray scattering (SAXS) setup to study biological macromolecules in solution. Our system consists of a microfocus X-ray tube delivering a highly collimated flux of 2.5 × 106 photons/s at a beam size of 1.2 × 1.2 mm2 at the collimation path exit and a maximum beam divergence of 0.16 mrad. The resulting observable scattering vectors q are in the range of 0.38 Å–1 down to 0.009 Å–1 in SAXS configuration and of 0.26 Å–1 up to 5.7 Å–1 in wide-angle X-ray scattering (WAXS) mode. To determine the capabilities of the instrument, we collected SAXS data on weakly scattering biological macromolecules including proteins and a nucleic acid sample with molecular weights varying from ~12 to 69 kDa and concentrations of 1.5–24 mg/ml. We report the measured scattering data display a high signal-to-noise ratio up to q-values of ~0.2 Å–1 allowing for an accurate structural characterization of the samples. Moreover, the in-house source data are of sufficient quality to perform ab initio 3D structure reconstructions that are in excellent agreement with the available crystallographic structures. In addition, measurements for the detergent decyl-maltoside show that the setup can be used to determine the size,more » shape, and interactions (as characterized by the second virial coefficient) of detergent micelles. This demonstrates that the use of a Mo-anode based in-house source is sufficient to determine basic geometric parameters and 3D shapes of biomolecules and presents a viable alternative to valuable beam time at third generation synchrotron sources.« less

Authors:
 [1];  [1];  [1];  [1];  [1];  [1]
  1. Ludwig Maximilian Univ., Munich (Germany). Dept. of Physics, Nanosystems Initiative Munich, and Center for Nanoscience
Publication Date:
Research Org.:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1238284
Resource Type:
Accepted Manuscript
Journal Name:
Review of Scientific Instruments
Additional Journal Information:
Journal Volume: 87; Journal Issue: 2; Journal ID: ISSN 0034-6748
Publisher:
American Institute of Physics (AIP)
Country of Publication:
United States
Language:
ENGLISH
Subject:
47 OTHER INSTRUMENTATION

Citation Formats

Bruetzel, Linda K., Fischer, Stefan, Salditt, Annalena, Sedlak, Steffen M., Nickel, Bert, and Lipfert, Jan. A Mo-anode-based in-house source for small-angle X-ray scattering measurements of biological macromolecules. United States: N. p., 2016. Web. doi:10.1063/1.4940936.
Bruetzel, Linda K., Fischer, Stefan, Salditt, Annalena, Sedlak, Steffen M., Nickel, Bert, & Lipfert, Jan. A Mo-anode-based in-house source for small-angle X-ray scattering measurements of biological macromolecules. United States. https://doi.org/10.1063/1.4940936
Bruetzel, Linda K., Fischer, Stefan, Salditt, Annalena, Sedlak, Steffen M., Nickel, Bert, and Lipfert, Jan. Mon . "A Mo-anode-based in-house source for small-angle X-ray scattering measurements of biological macromolecules". United States. https://doi.org/10.1063/1.4940936. https://www.osti.gov/servlets/purl/1238284.
@article{osti_1238284,
title = {A Mo-anode-based in-house source for small-angle X-ray scattering measurements of biological macromolecules},
author = {Bruetzel, Linda K. and Fischer, Stefan and Salditt, Annalena and Sedlak, Steffen M. and Nickel, Bert and Lipfert, Jan},
abstractNote = {We demonstrate the use of a molybdenum-anode-based in-house small-angle X-ray scattering (SAXS) setup to study biological macromolecules in solution. Our system consists of a microfocus X-ray tube delivering a highly collimated flux of 2.5 × 106 photons/s at a beam size of 1.2 × 1.2 mm2 at the collimation path exit and a maximum beam divergence of 0.16 mrad. The resulting observable scattering vectors q are in the range of 0.38 Å–1 down to 0.009 Å–1 in SAXS configuration and of 0.26 Å–1 up to 5.7 Å–1 in wide-angle X-ray scattering (WAXS) mode. To determine the capabilities of the instrument, we collected SAXS data on weakly scattering biological macromolecules including proteins and a nucleic acid sample with molecular weights varying from ~12 to 69 kDa and concentrations of 1.5–24 mg/ml. We report the measured scattering data display a high signal-to-noise ratio up to q-values of ~0.2 Å–1 allowing for an accurate structural characterization of the samples. Moreover, the in-house source data are of sufficient quality to perform ab initio 3D structure reconstructions that are in excellent agreement with the available crystallographic structures. In addition, measurements for the detergent decyl-maltoside show that the setup can be used to determine the size, shape, and interactions (as characterized by the second virial coefficient) of detergent micelles. This demonstrates that the use of a Mo-anode based in-house source is sufficient to determine basic geometric parameters and 3D shapes of biomolecules and presents a viable alternative to valuable beam time at third generation synchrotron sources.},
doi = {10.1063/1.4940936},
journal = {Review of Scientific Instruments},
number = 2,
volume = 87,
place = {United States},
year = {Mon Feb 08 00:00:00 EST 2016},
month = {Mon Feb 08 00:00:00 EST 2016}
}

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