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Open Dataset XS6ARGD0

Apoptosis-Inducing Factor (AIF) chimeric fusion with CHCHD4 N-terminal interaction domain

Abstract

This dataset captures a chimeric fusion of allostery mutant AIF-W196A (residues 104-613) and the AIF-interaction domain of CHCHD4 (residues 1-45). This chimeric protein was used to crystallize a regulatory complex of AIF and CHCHD4 – revealing how AIF regulates CHCHD4’s disulfide chaperone activity during mitochondrial protein import. SAXS data were used to establish the structural similarity between this fusion construct and the native NADH-activated AIF dimer. Corresponding X-ray crystal structure coordinates and SAXS data may be found at the Protein Data Bank with code 8VGY, and the SASBDB with code SASDV47, respectively. This dataset is described in further detail in Brosey CA, Shen R, and Tainer JA, The EMBO Journal, 2025.

Experimental description

The fusion construct AIF-W196A (104-613) – CHCHD4 (1-45) was expressed in bacteria and purified as described in Brosey et al, Structure, 2016. Purified AIF-CHCHD4 chimera was exchanged into SAXS buffer (25 mM HEPES, 150 mM NaCl, 2 mM TCEP, pH 7.5) by size-exclusion chromatography (Superdex 200 10/300) to ensure monodisperse protein. SAXS samples were prepared as a concentration series with SEC-matched buffers. The reported scattering curve was at 4.9 mg/mL. HT-SAXS data were collected by mail-in service at ALS beamline 12.3.1 (SIBYLS) at Lawrence Berkeley National Laboratory.

File description

The zip file contains time-resolved scattering curves (0.5 s, 29 frames, AIF-W196A-CHCHD4-45-chimera_####.dat) of AIF-CHCHD4-chimera, as well as a scattering curve averaged across all frames (AIF-W196A-CHCHD4-45-chimera_Average.dat).

Created

2024-03-30

Published

2025-03-04

Data collection technique

HT-SAXS

Journal DOI

https://doi.org/10.1038/s44318-024-00360-6

Source

Advanced Light Source

Beamline

SIBYLS BL12.3.1

Wavelength

1.27 Å

Sample to Detector Distance

2.0 m

Submitting Author

Chris Brosey

University of Texas MD Anderson Cancer Center

United States of America

[email protected]

Collaborators

Project Leader

John Tainer

[email protected]


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Complete Set of SAS Data Files

The complete SAS dataset is downloadable as a zip file.

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.

  • AIF-W196A-CHCHD4-45-chimera_Average.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.

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SAXS Similarity SAXS FrameSlice

Sample:

  • Macromolecule 1: AIF-W196A-CHCHD4-chimera
    • Sample Full Name: Apoptosis-Inducing Factor (AIF) W196A - CHCHD4 N-terminal chimera
    • Sample Type: Protein
    • Source Organism: Homo sapiens
    • Source Organism NCBI Taxonomy ID: 9606
    • Expression System: Escherichia coli
    • Expression NCBI Taxonomy ID: 562
    • Uniprot ID's: O95831, Q8N4Q1
    • Sequence or Chemical Formula:
    • MLTPEQKQKKAALSASEGEEVPQDKAPSHVPFLLIGGGTAAFAAARSIRARDPGARVLIVSEDPELPYMRPPLSKELWFSDDPNVTKTLRFKQANGKERSIYFQPPSFYVSAQDLPHIENGGVAVLTGKKVVQLDVRDNMVKLNDGSQITYEKCLIATGGTPRSLSAIDRAGAEVKSRTTLFRKIGDFRSLEKISREVKSITIIGGGFLGSELACALGRKARALGTEVIQLFPEKGNMGKILPEYLSNWTMEKVRREGVKVMPNAIVQSVGVSSGKLLIKLKDGRKVETDHIVAAVGLEPNVELAKTGGLEIDSDFGGFRVNAELQARSNIWVAGDAACFYDIKLGRRRVEHHDHAVVSGRLAGENMTGAAKPYWHQSMFWSDLGPDVGYEAIGLVDSSLPTVGVFAKATAQDNPKSATEQSGTGIRSESETESEASEITIPPSTPAVPQAPVQGEDYGKGVIFYLRDKVVVGIVLWNIFNRMPIARKIIKDGEQHEDLNEVAKLFNIHEDSGSGPGSGSMSYCRQEGKDRIIFVTKEDHETPSSAELVADDPNDPYEEHGLILPLEVLFQ