Complementary substrate specificity and distinct quaternary assembly of the Escherichia coli aerobic and anaerobic beta-oxidation trifunctional enzyme complexes.

Sah-Teli SK, Hynönen MJ, Schmitz W, Geraets JA, Seitsonen J, Pedersen JS, Butcher SJ, Wierenga RK, Venkatesan R, Biochem J (2019) Europe PMC

SASDEL9 – Escherichia coli aerobic fatty acid beta-oxidation trifunctional enzyme complex

Fatty acid oxidation complex subunit alpha
Fatty acid oxidation complex subunit alpha
3-ketoacyl-CoA thiolase FadA (beta subunit)
MWexperimental 233 kDa
MWexpected 244 kDa
VPorod 406 nm3
log I(s) 6.55×10-2 6.55×10-3 6.55×10-4 6.55×10-5
Fatty acid oxidation complex subunit alpha Fatty acid oxidation complex subunit alpha 3-ketoacyl-CoA thiolase FadA (beta subunit) small angle scattering data  s, nm-1
ln I(s)
Fatty acid oxidation complex subunit alpha Fatty acid oxidation complex subunit alpha 3-ketoacyl-CoA thiolase FadA (beta subunit) Guinier plot ln 6.56×10-2 Rg: 4.6 nm 0 (4.6 nm)-2 s2
(sRg)2I(s)/I(0)
Fatty acid oxidation complex subunit alpha Fatty acid oxidation complex subunit alpha 3-ketoacyl-CoA thiolase FadA (beta subunit) Kratky plot 1.104 0 3 sRg
p(r)
Fatty acid oxidation complex subunit alpha Fatty acid oxidation complex subunit alpha 3-ketoacyl-CoA thiolase FadA (beta subunit) pair distance distribution function Rg: 4.6 nm 0 Dmax: 16.0 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Fatty acid oxidation complex subunit alpha Fatty acid oxidation complex subunit alpha 3-ketoacyl-CoA thiolase FadA (beta subunit) DAMMIN model

log I(s)
 s, nm-1
Fatty acid oxidation complex subunit alpha Fatty acid oxidation complex subunit alpha 3-ketoacyl-CoA thiolase FadA (beta subunit) SWISSMODEL model

Synchrotron SAXS data from solutions of the E. coli aerobic fatty acid beta-oxidation trifunctional enzyme complex in 20 mM HEPES, 120 mM KCl, 2.5 mM DTT, pH 7.2 were measured at 20°C using size-exclusion chromatography SAXS (SEC-SAXS) on the B21 beam line at the Diamond Light Source (Oxfordshire, UK) using a Pilatus 2M detector at a sample-detector distance of 4.0 m and at a wavelength of λ = 0.1 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). The data were collected as 540 successive 3 second frames through the SEC elution profile (sample concentration = 6.20 mg/ml ). The SAXS data were measured from the sample and buffer from a Shodex KW403 column at flow rate of 0.16 mL/min. The sample-peak data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted and the individual subtracted data sets were scaled and averaged to generate the scattering profile displayed in this entry.

Fatty acid oxidation complex subunit alpha (EcTFE-alpha)
Mol. type   Protein
Organism   Escherichia coli (strain K12)
Olig. state   Monomer
Mon. MW   81.2 kDa
 
UniProt   P21177 (full sequence)
Sequence   FASTA
 
Fatty acid oxidation complex subunit alpha (EcTFE-alpha)
Mol. type   Protein
Organism   Escherichia coli (strain K12)
Olig. state   Monomer
Mon. MW   81.2 kDa
 
UniProt   P21177
Sequence   FASTA
 
3-ketoacyl-CoA thiolase FadA (beta subunit) (EcTFE-beta)
Mol. type   Protein
Organism   Escherichia coli (strain K12)
Olig. state   Dimer
Mon. MW   40.9 kDa
 
UniProt   P21151
Sequence   FASTA