Architecture and subunit dynamics of the mitochondrial TIM9·10·12 chaperone

Weinhäupl K, Wang Y, Hessel A, Brennich M, Lindorff-Larsen K, Schanda P, (2020) DOI

SASDH28 – Mitochondrial import inner membrane translocase TIM 9·10·12

Mitochondrial import inner membrane translocase subunit TIM12
Mitochondrial import inner membrane translocase subunit TIM9
Mitochondrial import inner membrane translocase subunit TIM10
MWexperimental 62 kDa
MWexpected 63 kDa
VPorod 103 nm3
log I(s) 5.96×101 5.96×100 5.96×10-1 5.96×10-2
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 small angle scattering data  s, nm-1
ln I(s)
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Guinier plot ln 5.96×101 Rg: 2.7 nm 0 (2.7 nm)-2 s2
(sRg)2I(s)/I(0)
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 Kratky plot 1.104 0 √3 sRg
p(r)
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 pair distance distribution function Rg: 2.7 nm 0 Dmax: 10 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 CUSTOM IN-HOUSE model
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 CUSTOM IN-HOUSE model
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 CUSTOM IN-HOUSE model
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 CUSTOM IN-HOUSE model
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 CUSTOM IN-HOUSE model
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 CUSTOM IN-HOUSE model
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 CUSTOM IN-HOUSE model
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 CUSTOM IN-HOUSE model
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 CUSTOM IN-HOUSE model
Mitochondrial import inner membrane translocase subunit TIM12 Mitochondrial import inner membrane translocase subunit TIM9 Mitochondrial import inner membrane translocase subunit TIM10 CUSTOM IN-HOUSE model
log I(s)
 s, nm-1
Synchrotron SAXS data from solutions of the mitochondrial import inner membrane translocase (TIM 9·10·12) in 20 mM MES pH 6.5, 50 mM NaCl 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 500.00 μl sample at 7 mg/ml was injected at a 0.50 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 20°C. 30 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.

Mitochondrial import inner membrane translocase subunit TIM12 (TIM12)
Mol. type   Protein
Organism   Saccharomyces cerevisiae
Olig. state   Monomer
Mon. MW   12.2 kDa
 
UniProt   P32830 (2-109)
Sequence   FASTA
 
Mitochondrial import inner membrane translocase subunit TIM9 (TIM9)
Mol. type   Protein
Organism   Saccharomyces cerevisiae
Olig. state   Dimer
Mon. MW   10.2 kDa
 
UniProt   O74700 (1-87)
Sequence   FASTA
 
Mitochondrial import inner membrane translocase subunit TIM10 (TIM10)
Mol. type   Protein
Organism   Saccharomyces cerevisiae
Olig. state   Trimer
Mon. MW   10.2 kDa
 
UniProt   P87108 (2-93)
Sequence   FASTA