Open Dataset XS97QA1S
Oxidized and X-ray-reduced Apoptosis-Inducing Factor with Allosteric Fragment Binders Examined by Time-resolved HT-SAXS
This dataset applies time-resolved HT-SAXS to identify and triage allosteric fragment binders of oxidized and X-ray-reduced Apoptosis-Inducing Factor (AIF). Fragment hits of AIF were identified by high-throughput differential scanning fluorescence (HT-DSF), then validated by DSF and microscale thermophoresis (MST). SAXS data with 300-ms framing were collected on 39 AIF-fragment complexes and analyzed at 0.3-s and 2.1-s exposures to determine fragment impact on oxidized and X-ray-reduced AIF redox states, respectively. These datasets revealed fragments capable of stimulating dimerization of X-ray-reduced AIF and triaged fragment binders into conformational chemotypes. This dataset is described in detail in a soon-to-be published manuscript.Experimental description
N-terminally truncated AIF (78-613) was expressed in bacteria and purified as described in Brosey et al, Structure, 2016. Purified AIF was exchanged into SAXS buffer by size-exclusion chromatography (Superdex 200 10/300) to ensure monodisperse protein. SAXS samples were prepared at 4 mg/mL with direct addition of DMSO or library fragments (10 mg/mL) to protein and SEC-matched buffers (Brosey et al, Methods in Enzymology, 2023). HT-SAXS data were collected by mail-in service at ALS beamline 12.3.1 (SIBYLS) at Lawrence Berkeley National Laboratory. **For deposition, this dataset was reprocessed and rescaled from the original image files to ensure correct intensity errors.**File description
Two zip files containing 0.3 s (oxidized) and 2.1 s (X-ray-reduced) SAXS curves for AIF-fragment complexes. File names are descriptive of DMSO or fragment identifier.
2023-04-14Data collection technique
Advanced Light SourceBeamline
1.27 ÅSample to Detector Distance
University of Texas MD Anderson Cancer Center
United States of America
Data Download Terms:
Complete Set of SAS Data Files
The complete SAS dataset is downloadable as a zip file.
- AIF-Allosteric-Fragments.zip Download
Individual SAS Data Files (total 1)
These are individual SAS data files that may be raw or processed (merged, etc.). These may or not be included in a zip file containing a larger dataset.
- 0.3s-DMSO.dat Download
Supplemental Data and Supporting Materials (total 0)
These data and materials may include X-ray crystal structure coordinates, multi-angle light scattering data, .etc. It may also include additional details or methods pertaining to the SAXS experiments, or the researcher's interpretations of the results.
Open SAXS data analysis sites in a new tab using these linksSAXS Similarity SAXS FrameSlice
- Macromolecule 1: AIF
- Sample Full Name: Apoptosis-inducing Factor
- Sample Type: Protein
- Source Organism: Homo sapiens
- Source Organism NCBI Taxonomy ID: 9606
- Expression System: Escherichia coli
- Expression NCBI Taxonomy ID: 562
- Uniprot ID: O95831
- Sequence or Chemical Formula: