Structural Basis of Membrane Protein Chaperoning through the Mitochondrial Intermembrane Space.

Weinhäupl K, Lindau C, Hessel A, Wang Y, Schütze C, Jores T, Melchionda L, Schönfisch B, Kalbacher H, Bersch B, Rapaport D, Brennich M, Lindorff-Larsen K, Wiedemann N, Schanda P, Cell 175(5):1365-1379.e25 (2018) Europe PMC

SASDEG2 – Mitochondrial import inner membrane translocase complex TIM9·10 in complex with a precursor (GDP/GTP carrier (Ggc1))

Mitochondrial import inner membrane translocase subunit TIM9
Mitochondrial import inner membrane translocase subunit TIM10
Mitochondrial GTP/GDP carrier protein 1
MWexperimental 160 kDa
MWexpected 156 kDa
VPorod 272 nm3
log I(s) 1.39×101 1.39×100 1.39×10-1 1.39×10-2
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 small angle scattering data  s, nm-1
ln I(s)
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 Guinier plot ln 1.39×101 Rg: 4.5 nm 0 (4.5 nm)-2 s2
(sRg)2I(s)/I(0)
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 Kratky plot 1.104 0 3 sRg
p(r)
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 pair distance distribution function Rg: 4.6 nm 0 Dmax: 16 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 DAMMIF model

log I(s)
 s, nm-1
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 CORAL model

log I(s)
 s, nm-1
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 GROMACS model
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 GROMACS model
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 GROMACS model
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 GROMACS model
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 GROMACS model
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 GROMACS model
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 GROMACS model
Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Mitochondrial GTP/GDP carrier protein 1 GROMACS model

Synchrotron SAXS data from solutions of the mitochondrial import inner membrane translocase complex (TIM9·10) in complex with a precursor (GDP/GTP carrier (Ggc1)) in 50mM Tris, 150mM NaCl, imidiazole, pH 7.4 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 (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Data was collected directly on the eluent from an Ni-NTA affinity chromatography column to ensure the isolation of a stable, non-aggregated complex. A solute concentration of approximately 2.00 mg/ml was measured at 20°C. 50 successive 1 second frames were collected through the elution peak. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Mitochondrial import inner membrane translocase subunit TIM9 (TIM9)
Mol. type   Protein
Organism   Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Olig. state   Hexamer
Mon. MW   10.2 kDa
 
UniProt   O74700
Sequence   FASTA
 
Mitochondrial import inner membrane translocase subunit TIM10 (TIM10)
Mol. type   Protein
Organism   Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Olig. state   Hexamer
Mon. MW   10.2 kDa
 
UniProt   P87108
Sequence   FASTA
 
Mitochondrial GTP/GDP carrier protein 1 (GGC1)
Mol. type   Protein
Organism   Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Olig. state   Monomer
Mon. MW   33.4 kDa
 
UniProt   P38988 (8-300)
Sequence   FASTA