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A General Small-Angle X-ray Scattering-Based Screening Protocol Validated for Protein–RNA Interactions

Title data

Chen, Po-Chia ; Masiewicz, Pawel ; Rybin, Vladimir ; Svergun, Dmitri ; Hennig, Janosch:
A General Small-Angle X-ray Scattering-Based Screening Protocol Validated for Protein–RNA Interactions.
In: ACS Combinatorial Science. Vol. 20 (2018) Issue 4 . - pp. 197-202.
ISSN 2156-8944
DOI: https://doi.org/10.1021/acscombsci.8b00007

Abstract in another language

We present a screening protocol utilizing small-angle X-ray scattering (SAXS) to obtain structural information on biomolecular interactions independent of prior knowledge, so as to complement affinity-based screening and provide leads for further exploration. This protocol categorizes ligand titrations by computing pairwise agreement between curves, and separately estimates affinities by quantifying complex formation as a departure from the linear sum properties of solution SAXS. The protocol is validated by sparse sequence search around the native poly uridine RNA motifs of the two-RRM domain Sex-lethal protein (Sxl). The screening of 35 RNA motifs between 4 to 10 nucleotides reveals a strong variation of resulting complexes, revealed to be preference-switching between 1:1 and 2:2 binding stoichiometries upon addition of structural modeling. Validation of select sequences in isothermal calorimetry and NMR titration retrieves domain-specific roles and function of a guanine anchor. These findings reinforce the suitability of SAXS as a complement in lead identification.

Further data

Item Type: Article in a journal
Refereed: Yes
Keywords: Structure-based screening; biomolecular interactions; RNA-binding proteins; small-angle X-ray scattering
Institutions of the University: Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biochemistry with an Emphasis on Biophysical Chemistry > Chair Biochemistry with an Emphasis on Biophysical Chemistry - Univ.-Prof. Dr. Janosch Hennig
Result of work at the UBT: No
DDC Subjects: 500 Science > 540 Chemistry
500 Science > 570 Life sciences, biology
Date Deposited: 07 Oct 2021 07:27
Last Modified: 07 Oct 2021 07:27
URI: https://eref.uni-bayreuth.de/id/eprint/67222