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Title: Absolute Intra-Molecular Distance Measurements with Ångström-Resolution using Anomalous Small-Angle X-ray Scattering

Accurate determination of molecular distances is fundamental to understanding the structure, dynamics, and conformational ensembles of biological macromolecules. Here we present a method to determine the full,distance,distribution between small (~7 Å) gold labels attached to macromolecules with very high-precision(≤1 Å) and on an absolute distance scale. Our method uses anomalous small-angle X-ray scattering close to a gold absorption edge to separate the gold-gold interference pattern from other scattering contributions. Results for 10-30 bp DNA constructs achieve excellent signal-to-noise and are in good agreement with previous results obtained by single-energy,SAXS measurements without requiring the preparation and measurement of single labeled and unlabeled samples. Finally, the use of small gold labels in combination with ASAXS read out provides an attractive approach to determining molecular distance distributions that will be applicable to a broad range of macromolecular systems.
Authors:
 [1] ;  [2] ;  [3] ;  [4] ;  [5] ;  [1]
  1. Ludwig Maximilian Univ., Munich (Germany). Nanosystems Initiative Munich, and Center for Nanoscience, Dept. of Physics
  2. Harvard Univ., Boston, MA (United States). Harvard Medical School, Dept. of Cell Biology
  3. Argonne National Lab. (ANL), Argonne, IL (United States). X-ray Science Division
  4. Stanford Univ., CA (United States). Dept. of Applied Physics and Physics
  5. Stanford Univ., CA (United States). Dept. of Biochemistry
Publication Date:
Grant/Contract Number:
AC02-06CH11357; DPOD000429
Type:
Accepted Manuscript
Journal Name:
Nano Letters
Additional Journal Information:
Journal Volume: 16; Journal Issue: 9
Research Org:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); National Institutes of Health (NIH)
Country of Publication:
United States
Language:
English
Subject:
47 OTHER INSTRUMENTATION; DNA; Intramolecular distances; SAXS; anomalous small-angle X-ray scattering; gold nanocrystals; molecular ruler
OSTI Identifier:
1373387

Zettl, Thomas, Mathew, Rebecca S., Seifert, Sönke, Doniach, Sebastian, Harbury, Pehr A.B., and Lipfert, Jan. Absolute Intra-Molecular Distance Measurements with Ångström-Resolution using Anomalous Small-Angle X-ray Scattering. United States: N. p., Web. doi:10.1021/acs.nanolett.6b01160.
Zettl, Thomas, Mathew, Rebecca S., Seifert, Sönke, Doniach, Sebastian, Harbury, Pehr A.B., & Lipfert, Jan. Absolute Intra-Molecular Distance Measurements with Ångström-Resolution using Anomalous Small-Angle X-ray Scattering. United States. doi:10.1021/acs.nanolett.6b01160.
Zettl, Thomas, Mathew, Rebecca S., Seifert, Sönke, Doniach, Sebastian, Harbury, Pehr A.B., and Lipfert, Jan. 2016. "Absolute Intra-Molecular Distance Measurements with Ångström-Resolution using Anomalous Small-Angle X-ray Scattering". United States. doi:10.1021/acs.nanolett.6b01160. https://www.osti.gov/servlets/purl/1373387.
@article{osti_1373387,
title = {Absolute Intra-Molecular Distance Measurements with Ångström-Resolution using Anomalous Small-Angle X-ray Scattering},
author = {Zettl, Thomas and Mathew, Rebecca S. and Seifert, Sönke and Doniach, Sebastian and Harbury, Pehr A.B. and Lipfert, Jan},
abstractNote = {Accurate determination of molecular distances is fundamental to understanding the structure, dynamics, and conformational ensembles of biological macromolecules. Here we present a method to determine the full,distance,distribution between small (~7 Å) gold labels attached to macromolecules with very high-precision(≤1 Å) and on an absolute distance scale. Our method uses anomalous small-angle X-ray scattering close to a gold absorption edge to separate the gold-gold interference pattern from other scattering contributions. Results for 10-30 bp DNA constructs achieve excellent signal-to-noise and are in good agreement with previous results obtained by single-energy,SAXS measurements without requiring the preparation and measurement of single labeled and unlabeled samples. Finally, the use of small gold labels in combination with ASAXS read out provides an attractive approach to determining molecular distance distributions that will be applicable to a broad range of macromolecular systems.},
doi = {10.1021/acs.nanolett.6b01160},
journal = {Nano Letters},
number = 9,
volume = 16,
place = {United States},
year = {2016},
month = {5}
}