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

SASDHX4 – Mitochondrial complex I assembly factor, ACAD9, chimeric construct where segment 445-482 has been replaced with segment 481-518 of VLCAD protein

Complex I assembly factor ACAD9-VLCAD chimera
MWI(0) 126 kDa
MWexpected 131 kDa
VPorod 217 nm3
log I(s) 1.42×102 1.42×101 1.42×100 1.42×10-1
Complex I assembly factor ACAD9-VLCAD chimera small angle scattering data  s, nm-1
ln I(s)
Complex I assembly factor ACAD9-VLCAD chimera Guinier plot ln 1.42×102 Rg: 3.9 nm 0 (3.9 nm)-2 s2
(sRg)2I(s)/I(0)
Complex I assembly factor ACAD9-VLCAD chimera Kratky plot 1.104 0 3 sRg
p(r)
Complex I assembly factor ACAD9-VLCAD chimera pair distance distribution function Rg: 3.7 nm 0 Dmax: 11.7 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Complex I assembly factor ACAD9-VLCAD chimera DAMFILT model
Complex I assembly factor ACAD9-VLCAD chimera DAMMIF model

Synchrotron SAXS data from solutions of an ACAD9 chimeric construct where segment 445-482 was replaced with segment 481-518 of VLCAD protein 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). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 250.00 μl sample at 9.1 mg/ml was injected at a 0.50 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 20°C. 100 successive 1 second frames were collected through the SEC elution peak. 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-VLCAD chimera (ACAD9/VLCAD-Chimera)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   65.4 kDa
Sequence   FASTA