Structural and functional insights underlying recognition of histidine phosphotransfer protein in fungal phosphorelay systems

Paredes-Martínez F, Eixerés L, Zamora-Caballero S, Casino P, Communications Biology 7(1) (2024) DOI

SASDSU5 – Histidine-phosphotransferase from Candida albicans

Phosphorelay intermediate protein YPD1
MWexperimental 15 kDa
MWexpected 19 kDa
VPorod 33 nm3
log I(s) 5.09×101 5.09×100 5.09×10-1 5.09×10-2
Phosphorelay intermediate protein YPD1 small angle scattering data  s, nm-1
ln I(s)
Phosphorelay intermediate protein YPD1 Guinier plot ln 5.09×101 Rg: 2.5 nm 0 (2.5 nm)-2 s2
Phosphorelay intermediate protein YPD1 Kratky plot 1.104 0 3 sRg
Phosphorelay intermediate protein YPD1 pair distance distribution function Rg: 2.5 nm 0 Dmax: 11.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Phosphorelay intermediate protein YPD1 SASREF model

Synchrotron SAXS data from solutions of Histidine-phosphotransferase in 50 mM Tris-HCl pH 8, 300 mM NaCl, were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus3 2M detector at a sample-detector distance of 2.8 m and at a wavelength of λ = 0.099 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 8 mg/ml was injected at a 0.07 ml/min flow rate onto a GE Superdex 75 Increase 3.2/300 column at 20°C. 1438 successive 2 second frames were collected across the entire SEC elution, where 76 sample frames were selected through the main SEC peak and used for averaging. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Poorly defined Guinier region.

Phosphorelay intermediate protein YPD1 (Cal_Ypd1)
Mol. type   Protein
Organism   Candida albicans (strain SC5314 / ATCC MYA-2876)
Olig. state   Monomer
Mon. MW   19.2 kDa
UniProt   Q59WC6 (12-184)
Sequence   FASTA