luxA Gene From Enhygromyxa salina Encodes a Functional Homodimeric Luciferase

Yudenko A, Bazhenov S, Aleksenko V, Goncharov I, Semenov O, Remeeva A, Nazarenko V, Kuznetsova E, Fomin V, Konopleva M, Al Ebrahim R, Sluchanko N, Ryzhykau Y, Semenov Y, Kuklin A, Manukhov I, Gushchin I, Proteins: Structure, Function, and Bioinformatics (2024) DOI

SASDQA7 – Homodimeric luciferase EsLuxA

Alkanal monooxygenase alpha chain
MWexperimental 99 kDa
MWexpected 84 kDa
VPorod 138 nm3
log I(s) 2.49×10-1 2.49×10-2 2.49×10-3 2.49×10-4
Alkanal monooxygenase alpha chain small angle scattering data  s, nm-1
ln I(s)
Alkanal monooxygenase alpha chain Guinier plot ln 2.49×10-1 Rg: 3.3 nm 0 (3.3 nm)-2 s2
(sRg)2I(s)/I(0)
Alkanal monooxygenase alpha chain Kratky plot 1.104 0 √3 sRg
p(r)
Alkanal monooxygenase alpha chain pair distance distribution function Rg: 3.3 nm 0 Dmax: 12.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Alkanal monooxygenase alpha chain ALPHAFOLD model
Alkanal monooxygenase alpha chain ALPHAFOLD model
Alkanal monooxygenase alpha chain ALPHAFOLD model
SAXS data from solutions of luciferase EsLuxA in 150 mM NaCl, 25 mM Tris, pH 8 were collected using a Rigaku MicroMax 007-HF instrument at the Moscow Institute of Physics and Technology (MIPT; Dolgoprudny, Russian Federation) equipped with a Multiwire gas-filled ASM DTR Triton 200 detector at a sample-detector distance of 2 m and at a wavelength of λ = 0.1504 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 3.20 mg/ml was measured at 20°C. Two successive 4200 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.

The protein of study is a luciferase LuxA from Enhygromyxa salina ((UniProt ID A0A2S9XZH0). According to the OLIGOMER approximation, dimers are the predominant oligomeric state (82%) of the EsLuxA. The rest of the protein can be described as tetramers (dimers of dimers), which account for 18%, while the proportion of monomers is negligible.

Alkanal monooxygenase alpha chain (LuxA)
Mol. type   Protein
Organism   Enhygromyxa salina
Olig. state   Dimer
Mon. MW   42.2 kDa
 
UniProt   A0A2S9XZH0 (1-363)
Sequence   FASTA