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Small angle neutron scattering for structural biology of challenging biomacromoleular complexes

Small angle neutron scattering (SANS) provides unique insight into biomacromolecular complexes by combining solvent contrast variation (H2O:D2O exchange) with either natural contrast between different classes of biomolecules (proteins, RNA/DNA, lipids/detergents) and/or by applying artificial contrast, i.e. deuteration of specific biomolecules. I will present several biological projects where SANS has played a crucial role by providing unique restraints for structural refinement and interpretation, complementary to NMR, SAXS, crystallography and EM.

1. A first example, I will show how distance and shape restraints from SANS have helped to improve the uniqueness of structural models for two multi-protein-RNA complexes, in combination with NMR restraints and building blocks from crystallography [1, 2]. In a second example, the stoichiometry and internal topology of a highly symmetric, hetero-dodecameric aminopeptidase enzyme complex is revealed by SANS, and conclusions on the assembling process can be drawn in combination with EM data [3]. Other examples will include structural studies of two solubilized membrane proteins and their detergent belts [4, 5], a time-resolved study of the active unfolding of GFP by an unfoldase complex [6], and insight into the hydration shell properties of proteins [7].

2. I will present two recent developments that will improve the quality and accuracy of SANS data: one the one hand, segmental deuteration which allows to access more detailed structural information for the important class of multi-domain proteins [8], and on the other hand size-exclusion chromatography (SEC) coupled to SANS, which improves sample quality, and in particular monodispersity [9].

References:

[1] Lapinaite et al. (2013) Nature502(7472), 519-523.
[2] Hennig et al. (2014) Nature515(7526), 287-290.
[3] Appolaire et al. (2014)Acta Cryst. D70, 2983-2993.
[4] Gabel et al. (2014) Biophys. J.107(1), 185-196.
[5] Dias-Mirandela et al. (2018) J. Phys. Chem. Lett.9(14), 3910-3914.
[6] Ibrahim et al. (2017) Sci. Rep.7, 40948.
[7] Kim et al. (2016) Biophys. J.110(10), 2185-2194.
[8] Sonntag et al. (2017) Angew. Chem.56(32), 9322-9325.
[9] Jordan et al. (2016) J. Appl. Crystallogr.49(Pt 6), 2015-2020.

Seminar: Neutronen in Forschung und Industrie

Datum12.11.2018
Uhrzeit14:30 - 15:30 Uhr
OrtGarching / Physik Department
RaumHS 3
SprecherDr. Frank Gabel, IBS & ILL, Grenoble, France
Veranstalter

TUM / MLZ


Kontakt

Ansprechpartner

Dr. Markos Skoulatos und Dr.Alexandros Koutsioumpas

E-Mail

Markos.Skoulatos@frm2.tum.de

URL

https://indico.frm2.tum.de/e/159

MLZ ist eine Kooperation aus:

Technische Universität München> Technische Universität MünchenHelmholtz-Zentrum Hereon> Helmholtz-Zentrum Hereon
Forschungszentrum Jülich> Forschungszentrum Jülich

MLZ ist Mitglied in:

LENS> LENSERF-AISBL> ERF-AISBL

MLZ in den sozialen Medien: