Assembly of the mitochondrial Complex I assembly complex suggests a regulatory role for deflavination.

Giachin G, Jessop M, Bouverot R, Acajjaoui S, Saidi M, Chretien A, Bacia-Verloop M, Signor L, Mas PJ, Favier A, Borel Meneroud E, Hons M, Hart DJ, Kandiah E, Boeri Erba E, Buisson A, Leonard G, Gutsche I, Soler-Lopez M, Angew Chem Int Ed Engl (2020) Europe PMC

SASDHZ4 – The C-terminal domain of ESCIT bound to a dimer of the complex I assembly factor ACAD9

Complex I assembly factor ACAD9, mitochondrial
Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial
MWI(0) 280 kDa
MWexpected 219 kDa
VPorod 683 nm3
log I(s) 2.40×102 2.40×101 2.40×100 2.40×10-1
Complex I assembly factor ACAD9, mitochondrial Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial small angle scattering data  s, nm-1
ln I(s)
Complex I assembly factor ACAD9, mitochondrial Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial Guinier plot ln 2.41×102 Rg: 6.0 nm 0 (6.0 nm)-2 s2
(sRg)2I(s)/I(0)
Complex I assembly factor ACAD9, mitochondrial Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial Kratky plot 1.104 0 √3 sRg
p(r)
Complex I assembly factor ACAD9, mitochondrial Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial pair distance distribution function Rg: 5.8 nm 0 Dmax: 16.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Complex I assembly factor ACAD9, mitochondrial Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial GASBOR model
Synchrotron SAXS data from solutions of the C-terminal domain of ESCIT bound to a dimer of the complex I assembly factor ACAD9 in 25 mM HEPES, 250 mM NaCl, pH 7.5 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.9 m and at a wavelength of λ = 0.099 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 0.8 and 2 mg/ml were measured at 20°C. 10 successive 0.500 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Complex formed by the interaction between ECSIT C-terminus and dimeric ACAD9. Native-MS experiments suggest that the complex has mainly a 2:4 ACAD9:ECSIT stoichiometric ratio. Sample was immediatly measured after SEC purification.

Complex I assembly factor ACAD9, mitochondrial (ACAD9)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   66.0 kDa
 
UniProt   Q9H845 (38-621)
Sequence   FASTA
 
Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial (ECSIT)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Tetramer
Mon. MW   21.9 kDa
 
UniProt   Q9BQ95 (249-431)
Sequence   FASTA