The Methionine-Rich Loop of Multicopper Oxidase McoA follows Open-To-Close Transitions with a Role in Enzyme Catalysis

Borges P, Brissos V, Hernandez G, Masgrau L, Lucas M, Monza E, Frazão C, Cordeiro T, Martins L, ACS Catalysis (2020) DOI

SASDFW7 – Aquifex aeolicus McoA metaloxidase deletion mutant ∆328-352 (MCoA∆328-352)

Aquifex aeolicus McoA metaloxidase ∆328-352 (MCoA∆328-352)
MWexperimental 48 kDa
MWexpected 53 kDa
VPorod 77 nm3
log I(s) 2.27×101 2.27×100 2.27×10-1 2.27×10-2
Aquifex aeolicus McoA metaloxidase ∆328-352 (MCoA∆328-352) small angle scattering data  s, nm-1
ln I(s)
Aquifex aeolicus McoA metaloxidase ∆328-352 (MCoA∆328-352) Guinier plot ln 2.27×101 Rg: 2.3 nm 0 (2.3 nm)-2 s2
(sRg)2I(s)/I(0)
Aquifex aeolicus McoA metaloxidase ∆328-352 (MCoA∆328-352) Kratky plot 1.104 0 √3 sRg
p(r)
Aquifex aeolicus McoA metaloxidase ∆328-352 (MCoA∆328-352) pair distance distribution function Rg: 2.2 nm 0 Dmax: 6.9 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Aquifex aeolicus McoA metaloxidase ∆328-352 (MCoA∆328-352) DAMFILT model
log I(s)
 s, nm-1
Aquifex aeolicus McoA metaloxidase ∆328-352 (MCoA∆328-352) OTHER model
Synchrotron SAXS data from solutions of MCoA∆328-352 in 50 mM Tris-HCl, 150 mM NaCl, 2 mM TCEP, 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 = 0.09919 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 45.00 μl sample at 12 mg/ml was injected at a 0.10 ml/min flow rate onto a GE Superdex 200 Increase 3.2/300 column at 20°C. 3000 successive 1 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 from the sample elution peak frames.

Aquifex aeolicus McoA metaloxidase ∆328-352 (MCoA∆328-352) (MCoA∆328-352)
Mol. type   Protein
Organism   Aquifex aeolicus
Olig. state   Monomer
Mon. MW   52.9 kDa
 
UniProt   O67206 (44-527)
Sequence   FASTA