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- Title
Combined small angle X-ray solution scattering with atomic force microscopy for characterizing radiation damage on biological macromolecules.
- Authors
Costa, Luca; Andriatis, Alexander; Brennich, Martha; Teulon, Jean-Marie; Chen, Shu-wen W.; Pellequer, Jean-Luc; Round, Adam
- Abstract
Background: Synchrotron radiation facilities are pillars of modern structural biology. Small-Angle X-ray scattering performed at synchrotron sources is often used to characterize the shape of biological macromolecules. A major challenge with high-energy X-ray beam on such macromolecules is the perturbation of sample due to radiation damage. Results: By employing atomic force microscopy, another common technique to determine the shape of biological macromolecules when deposited on flat substrates, we present a protocol to evaluate and characterize consequences of radiation damage. It requires the acquisition of images of irradiated samples at the single molecule level in a timely manner while using minimal amounts of protein. The protocol has been tested on two different molecular systems: a large globular tetremeric enzyme (β-Amylase) and a rod-shape plant virus (tobacco mosaic virus). Radiation damage on the globular enzyme leads to an apparent increase in molecular sizes whereas the effect on the long virus is a breakage into smaller pieces resulting in a decrease of the average long-axis radius. Conclusions: These results show that radiation damage can appear in different forms and strongly support the need to check the effect of radiation damage at synchrotron sources using the presented protocol.
- Subjects
SYNCHROTRON radiation; X-ray scattering; MACROMOLECULES; RADIATION damage; RADIOBIOLOGY
- Publication
BMC Structural Biology, 2016, Vol 16, p1
- ISSN
1472-6807
- Publication type
Article
- DOI
10.1186/s12900-016-0068-2