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

SASDJ34 – Human Complex I assembly factor ACAD9 dimer

Complex I assembly factor ACAD9, mitochondrial

MW^experimental	138	kDa
MW^expected	130	kDa
V^Porod	213	nm³

log I(s) 1.50×10² 1.50×10¹ 1.50×10⁰ 1.50×10^-1

Complex I assembly factor ACAD9, mitochondrial small angle scattering data

s, nm^-1

Log plot Log-Log plot

ln I(s)

Complex I assembly factor ACAD9, mitochondrial Guinier plot

ln 1.50×10² R_g: 3.5 nm 0 (3.5 nm)^-2 s²

(sR_g)²I(s)/I(0)

Complex I assembly factor ACAD9, mitochondrial Kratky plot

1.104 0 √3 sR_g

p(r)	R_g: 3.5 nm 0 D_max: 13 nm

Data validation

Experimental data range

p(r) fit to data

Metric

Value

s_min D_max / π

0.4

N_Shannon

Worse

Better

Metric

Value

p_cormap

χ²

0.004
1.131

Worse

Better

Fits and models

log I(s)	s, nm^-1
+3 Δ/σ -3	s, nm^-1

Complex I assembly factor ACAD9, mitochondrial CUSTOM IN-HOUSE model

log I(s)	s, nm^-1
+3 Δ/σ -3	s, nm^-1

Complex I assembly factor ACAD9, mitochondrial DAMMIF model

log I(s)	s, nm^-1
+3 Δ/σ -3	s, nm^-1

Complex I assembly factor ACAD9, mitochondrial DAMFILT model

Synchrotron SAXS data from solutions of ACAD9 dimer 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 7.1 mg/ml was injected at a 0.75 ml/min flow rate onto a GE Superose 6 Increase 10/300 column at 22°C. 568 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		Dimer
Mon. MW		64.8 kDa

UniProt		Q9H845 (38-621)
Sequence		FASTA