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

SASDFY7 – Aquifex aeolicus McoA metaloxidase

Aquifex aeolicus McoA metaloxidase
MWexperimental 50 kDa
MWexpected 55 kDa
VPorod 79 nm3
log I(s) 7.71×10-2 7.71×10-3 7.71×10-4 7.71×10-5
Aquifex aeolicus McoA metaloxidase small angle scattering data  s, nm-1
ln I(s)
Aquifex aeolicus McoA metaloxidase Guinier plot ln 7.72×10-2 Rg: 2.3 nm 0 (2.3 nm)-2 s2
(sRg)2I(s)/I(0)
Aquifex aeolicus McoA metaloxidase Kratky plot 1.104 0 √3 sRg
p(r)
Aquifex aeolicus McoA metaloxidase pair distance distribution function Rg: 2.3 nm 0 Dmax: 7.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Aquifex aeolicus McoA metaloxidase DAMFILT model
log I(s)
 s, nm-1
Aquifex aeolicus McoA metaloxidase OTHER model
Synchrotron SAXS data from solutions of McoA in 50 mM Tris-HCl, 150 mM NaCl, 2 mM TCEP, pH 7.5 were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Pilatus 2M detector at a sample-detector distance of 2.7 m and at a wavelength of λ = 0.0914 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 50.00 μl sample at 20 mg/ml was injected at a 0.16 ml/min flow rate onto a Shodex KW403 column at 20°C. 915 successive 3 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 (McoA)
Mol. type   Protein
Organism   Aquifex aeolicus
Olig. state   Monomer
Mon. MW   55.5 kDa
 
UniProt   O67206 (44-527)
Sequence   FASTA