Molecular mechanism of interactions between ACAD9 and binding partners in mitochondrial respiratory complex I assembly

Xia C, Lou B, Fu Z, Mohsen A, Shen A, Vockley J, Kim J, iScience 24(10):103153 (2021) DOI

SASDJ54 – Human Complex I assembly factor ACAD9 complexed with ECSIT and SMT3-NDUFAF1

Complex I assembly factor ACAD9, mitochondrial
Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial
Complex I intermediate-associated protein 30, mitochondrial
MWexperimental 715 kDa
MWexpected 634 kDa
VPorod 1340 nm3
log I(s) 1.14×102 1.14×101 1.14×100 1.14×10-1
Complex I assembly factor ACAD9, mitochondrial Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial Complex I intermediate-associated protein 30, 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 Complex I intermediate-associated protein 30, mitochondrial Guinier plot ln 1.15×102 Rg: 6.6 nm 0 (6.6 nm)-2 s2
(sRg)2I(s)/I(0)
Complex I assembly factor ACAD9, mitochondrial Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial Complex I intermediate-associated protein 30, mitochondrial Kratky plot 1.104 0 √3 sRg
p(r)
Complex I assembly factor ACAD9, mitochondrial Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial Complex I intermediate-associated protein 30, mitochondrial pair distance distribution function Rg: 6.8 nm 0 Dmax: 21.6 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 Complex I intermediate-associated protein 30, mitochondrial DAMMIF model
log I(s)
 s, nm-1
Complex I assembly factor ACAD9, mitochondrial Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial Complex I intermediate-associated protein 30, mitochondrial DAMFILT model
Synchrotron SAXS data from solutions of ACAD9 complexed with ECSIT and SMT3-NDUFAF1 in 25mM Tris-HCl, 150mM NaCl, 0.1mM DTT, and 5% glycerol, pH 8 were collected on the BioCAT 18ID beam line at the Advanced Photon Source (APS), Argonne National Laboratory (Lemont, IL, USA) using a Pilatus3 X 1M detector at a sample-detector distance of 3.5 m and at a wavelength of λ = 0.1033 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 250.00 μl sample at 1 mg/ml was injected at a 0.75 ml/min flow rate onto a GE Superose 6 Increase 10/300 column at 22°C. 550 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.

Complex I assembly factor ACAD9, mitochondrial (ACAD9)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Tetramer
Mon. MW   64.8 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   46.1 kDa
 
UniProt   Q9BQ95 (49-431)
Sequence   FASTA
 
Complex I intermediate-associated protein 30, mitochondrial (NDUFAF1)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Tetramer
Mon. MW   47.5 kDa
 
UniProt   Q9Y375 (25-327)
Sequence   FASTA