Determination of absolute intramolecular distances in proteins using anomalous X-ray scattering interferometry.

Autor: Stubhan S; Department of Physics and Center for NanoScience, LMU Munich, Amalienstr. 54, 80799 Munich, Germany.; Soft Condensed Matter and Biophysics, Department of Physics and Debye Institute for Nanomaterials Science, Utrecht University, Princetonplein 1, 3584 CC Utrecht, The Netherlands. J.Lipfert@uu.nl., Baptist AV; Department of Physics and Center for NanoScience, LMU Munich, Amalienstr. 54, 80799 Munich, Germany.; Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany., Körösy C; Soft Condensed Matter and Biophysics, Department of Physics and Debye Institute for Nanomaterials Science, Utrecht University, Princetonplein 1, 3584 CC Utrecht, The Netherlands. J.Lipfert@uu.nl., Narducci A; Physical and Synthetic Biology, Faculty of Biology, LMU Munich, Großhadernerstr. 2-4, 82152 Planegg-Martinsried, Germany. cordes@bio.lmu.de., Moya Muñoz GG; Physical and Synthetic Biology, Faculty of Biology, LMU Munich, Großhadernerstr. 2-4, 82152 Planegg-Martinsried, Germany. cordes@bio.lmu.de., Wendler N; Physical and Synthetic Biology, Faculty of Biology, LMU Munich, Großhadernerstr. 2-4, 82152 Planegg-Martinsried, Germany. cordes@bio.lmu.de., Lak A; Department of Physics and Center for NanoScience, LMU Munich, Amalienstr. 54, 80799 Munich, Germany., Sztucki M; ESRF, 71 Avenue des Martyrs, 38043 Grenoble, France., Cordes T; Physical and Synthetic Biology, Faculty of Biology, LMU Munich, Großhadernerstr. 2-4, 82152 Planegg-Martinsried, Germany. cordes@bio.lmu.de.; Biophysical Chemistry, Faculty of Chemistry and Chemical Biology, Technische Universität Dortmund, Otto-Hahn-Str. 4a, 44227 Dortmund, Germany., Lipfert J; Department of Physics and Center for NanoScience, LMU Munich, Amalienstr. 54, 80799 Munich, Germany.; Soft Condensed Matter and Biophysics, Department of Physics and Debye Institute for Nanomaterials Science, Utrecht University, Princetonplein 1, 3584 CC Utrecht, The Netherlands. J.Lipfert@uu.nl.; Institute for Physics, Augsburg University, Universitätsstrasse 1, 86159 Augsburg, Germany.
Jazyk: angličtina
Zdroj: Nanoscale [Nanoscale] 2024 Dec 18. Date of Electronic Publication: 2024 Dec 18.
DOI: 10.1039/d4nr03375b
Abstrakt: Biomolecular structures are typically determined using frozen or crystalline samples. Measurement of intramolecular distances in solution can provide additional insights into conformational heterogeneity and dynamics of biological macromolecules and their complexes. The established molecular ruler techniques used for this (NMR, FRET, and EPR) are, however, limited in their dynamic range and require model assumptions to determine absolute distance or distance distributions. Here, we introduce anomalous X-ray scattering interferometry (AXSI) for intramolecular distance measurements in proteins, which are labeled at two sites with small gold nanoparticles of 0.7 nm radius. We apply AXSI to two different cysteine-variants of maltose binding protein in the presence and absence of its ligand maltose and find distances in quantitative agreement with single-molecule FRET experiments. Our study shows that AXSI enables determination of intramolecular distance distributions under virtually arbitrary solution conditions and we anticipate its broad use to characterize protein conformational ensembles and dynamics.
Databáze: MEDLINE
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