Online data analysis at the ESRF bioSAXS beamline, BM29

Brennich, Martha; Kieffer, Jérôme; Bonamis, Guillaume; Antolinos, Alejandro De María; Hutin, Stephanie; Pernot, Pétra; Round, Adam

doi:10.1107/s1600576715024462

Cited by 82 publications

(75 citation statements)

References 35 publications

(42 reference statements)

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“…The available q -range () was 0.025 nm −1 to 4.8 nm −1 . Data reduction was done using the standard tools at BM29 yielding the 1D subtracted curves and initial processing to give feedback to the experiment [39, 40]. Extrapolation to zero concentration and determination of the model independent parameters ( R g (radius of gyration), I 0 (Intensity at q = 0), molecular mass, etc.)…”

Section: Methodsmentioning

confidence: 99%

Combined small angle X-ray solution scattering with atomic force microscopy for characterizing radiation damage on biological macromolecules

et al. 2016

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BackgroundSynchrotron 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.ResultsBy 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.ConclusionsThese 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.

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Section: Methodsmentioning

confidence: 99%

Combined small angle X-ray solution scattering with atomic force microscopy for characterizing radiation damage on biological macromolecules

et al. 2016

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“…55 (automated sample changer) or 100 (SEC) µl volumes were exposed to X-rays for 10 individual frames, each 1 second in duration. Individual frames were processed with the automatic processing pipeline 52 , to yield individual radially-averaged curves of normalized intensity versus scattering angle s = 4πsinθ/λ.…”

Section: Saxs Characterizationmentioning

confidence: 99%

Noncovalent PEGylation via Lectin–Glycopolymer Interactions

et al. 2016

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The full-text may be used and/or reproduced, and given to third parties in any format or medium, without prior permission or charge, for personal research or study, educational, or not-for-prot purposes provided that:• a full bibliographic reference is made to the original source • a link is made to the metadata record in DRO • the full-text is not changed in any way The full-text must not be sold in any format or medium without the formal permission of the copyright holders.Please consult the full DRO policy for further details. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 2 AbstractPEGylation, the covalent modification of proteins with polyethylene glycol, is an abundantly used technique to improve the pharmacokinetics of therapeutic proteins. The drawback with this methodology is that the covalently attached PEG can impede the biological activity (e.g. reduced receptor-binding capacity). Protein therapeutics with "disposable" PEG modifiers have potential advantages over the current technology. Here, we show that a protein-polymer "Medusa complex" is formed by the combination of a hexavalent lectin with a glycopolymer. Using NMR spectroscopy, small angle X-ray scattering (SAXS), size exclusion chromatography and native gel electrophoresis it was demonstrated that the fucose-binding lectin RSL and a fucose-capped polyethylene glycol (Fuc-PEG) form a multimeric assembly. All of the experimental methods provided evidence of noncovalent PEGylation with a concomitant increase in molecular mass and hydrodynamic radius. The affinity of the protein-polymer complex was determined by ITCand competition experiments to be in the µM range, suggesting that such systems have potential biomedical applications.

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“…The morphology of assembled DB921 nanotubes in NaCl was studied using solution‐state small‐angle X‐ray scattering (SAXS) and neutron scattering (SANS), performed on the BM29, ID02 beamlines at the ESRF, and instrument D11 at the ILL, respectively . Complementary cryo‐transmission electron microscopy (TEM) of the assembled nanotubes was also performed to provide direct visualization of the nanotubes.…”

Section: Resultsmentioning

confidence: 99%

Hierarchical Nanotube Self‐Assembly of DNA Minor Groove‐Binding Ligand DB921 via Alkali Halide Triggering

Mizuta

Devos

Narayanan

et al. 2019

Macromolecular Symposia

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A systematic study into a novel self‐assembling system is presented whereby the small molecule DB921 forms nearly monodisperse nanotubes upon addition of a suitable salt. Small‐angle X‐ray scattering and negative stain transmission electron microscopy are employed as the principal characterization methods to study both the fundamental assembly processes and reaction kinetics. The assembly is found to be hierarchical, culminating with helical winding of intermediate ribbon structures into nanotubes. The driving force for this process is likely the electrostatic interaction between DB921 and salt anions; this aspect can be used to control nanotube assembly and morphology.

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Online data analysis at the ESRF bioSAXS beamline, BM29

Abstract: International audienc

Cited by 82 publications

References 35 publications

Combined small angle X-ray solution scattering with atomic force microscopy for characterizing radiation damage on biological macromolecules

Combined small angle X-ray solution scattering with atomic force microscopy for characterizing radiation damage on biological macromolecules

Noncovalent PEGylation via Lectin–Glycopolymer Interactions

Hierarchical Nanotube Self‐Assembly of DNA Minor Groove‐Binding Ligand DB921 via Alkali Halide Triggering

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