ArticlePloS one2013
Tools for the quantitative analysis of sedimentation boundaries detected by fluorescence optical analytical ultracentrifugation.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
25 citing papers in PubMed.
- Phospholipid-driven conformational switching of HCV NS5A links protein folding to replication membrane remodeling.Science advances · 2026Article
- Analysis of Giant-Shell CdSe/CdS Quantum Dots via Analytical Ultracentrifugation Combined with Spectrally Resolved Photoluminescence.Small methods · 2026Article
- Development of an advanced multiwavelength emission detector for the analytical ultracentrifuge.Nanoscale advances · 2024Article
- Insight into the Structural Basis for Dual Nucleic Acid-Recognition by the Scaffold Attachment Factor B2 Protein.International journal of molecular sciences · 2023Article
- Unidirectional mannitol synthesis ofProceedings of the National Academy of Sciences of the United States of America · 2022Article
- Analytical Ultracentrifugation (AUC): An Overview of the Application of Fluorescence and Absorbance AUC to the Study of Biological Macromolecules.Current protocols in molecular biology · 2020Article
- A multiwavelength emission detector for analytical ultracentrifugation.Nanoscale advances · 2019Article
- Viroid research and its significance for RNA technology and basic biochemistry.Nucleic acids research · 2018Review
- Sedimentation velocity analytical ultracentrifugation for characterization of therapeutic antibodies.Biophysical reviews · 2018Review
- Article
- Microtubule cross-linking activity of She1 ensures spindle stability for spindle positioning.The Journal of cell biology · 2017Article
- Use of fluorescence-detected sedimentation velocity to study high-affinity protein interactions.Nature protocols · 2017Article
- Sedimentation of Reversibly Interacting Macromolecules with Changes in Fluorescence Quantum Yield.Biophysical journal · 2017Article
- EGFP oligomers as natural fluorescence and hydrodynamic standards.Scientific reports · 2016Article
- Sedimentation coefficient distributions of large particles.The Analyst · 2016Article
- Article
- Higher-order oligomerization promotes localization of SPOP to liquid nuclear speckles.The EMBO journal · 2016Article
- Tubulin Dimer Reversible Dissociation: AFFINITY, KINETICS, AND DEMONSTRATION OF A STABLE MONOMER.The Journal of biological chemistry · 2016Article
- 3D-Printing for Analytical Ultracentrifugation.PloS one · 2016Article
- Two-photon-like microscopy with orders-of-magnitude lower illumination intensity via two-step fluorescence.Nature communications · 2015Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
Fluorescence optical detection in sedimentation velocity analytical ultracentrifugation allows the study of macromolecules at nanomolar concentrations and below. This has significant promise, for example, for the study of systems of high-affinity protein interactions. Here we describe adaptations of the direct boundary modeling analysis approach implemented in the software SEDFIT that were developed to accommodate unique characteristics of the confocal fluorescence detection system. These include spatial gradients of signal intensity due to scanner movements out of the plane of rotation, temporal intensity drifts due to instability of the laser and fluorophores, and masking of the finite excitation and detection cone by the sample holder. In an extensive series of experiments with enhanced green fluorescent protein ranging from low nanomolar to low micromolar concentrations, we show that the experimental data provide sufficient information to determine the parameters required for first-order approximation of the impact of these effects on the recorded data. Systematic deviations of fluorescence optical sedimentation velocity data analyzed using conventional sedimentation models developed for absorbance and interference optics are largely removed after these adaptations, resulting in excellent fits that highlight the high precision of fluorescence sedimentation velocity data, thus allowing a more detailed quantitative interpretation of the signal boundaries that is otherwise not possible for this system.
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Registered trials
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