The disordered C-terminal tail of fungal LPMOs from phytopathogens mediates protein dimerization and impacts plant penetration.

Tamburrini KC, Kodama S, Grisel S, Haon M, Nishiuchi T, Bissaro B, Kubo Y, Longhi S, Berrin JG, Proc Natl Acad Sci U S A 121(13):e2319998121 (2024) Europe PMC

SASDUH3 – Colletotrichum orbiculare Auxiliary Activity Family 9A Lytic Polysaccharide Monooxygenase (AA9A LPMO) dimer

Endoglucanase-7
MWexperimental 53 kDa
MWexpected 60 kDa
VPorod 83 nm3
log I(s) 4.78×10-2 4.78×10-3 4.78×10-4 4.78×10-5
Endoglucanase-7 small angle scattering data  s, nm-1
ln I(s)
Endoglucanase-7 Guinier plot ln 4.79×10-2 Rg: 4.8 nm 0 (4.8 nm)-2 s2
(sRg)2I(s)/I(0)
Endoglucanase-7 Kratky plot 1.104 0 √3 sRg
p(r)
Endoglucanase-7 pair distance distribution function Rg: 5.2 nm 0 Dmax: 21.7 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Colletotrichum orbiculare Auxiliary Activity Family 9A Lytic Polysaccharide Monooxygenase (AA9A LPMO) dimer Rg histogram Rg, nm
Endoglucanase-7 EOM/RANCH model
Endoglucanase-7 EOM/RANCH model
Endoglucanase-7 EOM/RANCH model
Endoglucanase-7 EOM/RANCH model
log I(s)
 s, nm-1
Endoglucanase-7 GASBOR model
Synchrotron SAXS data from solutions of AA9 LPMO in 50 mM MES, 150 mM NaCl, pH 6.5 were collected on the SWING beam line at SOLEIL (Saint-Aubin, France) using a Eiger 4M detector at a sample-detector distance of 2 m and at a wavelength of λ = 0.1033 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 5.4 mg/ml was injected at a 0.30 ml/min flow rate onto a Agilent AdvanceBio SEC 300Å, 4.6 x 150 mm column at 20°C. 720 successive 0.990 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.

CoAA9A was heterologously expressed in Pichia pastoris. The sample was N-deglycosylated before recording SAXS data, however it was not possible to remove O-glycosylation. The difference between the experimental molecular weight and the expected molecular weight is due to O-glycosylation.

Endoglucanase-7 (AA9A LPMO)
Mol. type   Protein
Organism   Colletotrichum orbiculare (strain 104-T / ATCC 96160 / CBS 514.97 / LARS 414 / MAFF 240422)
Olig. state   Dimer
Mon. MW   29.9 kDa
 
UniProt   N4UN01 (21-303)
Sequence   FASTA