Reaction hijacking of tyrosine tRNA synthetase as a new whole-of-life-cycle antimalarial strategy

Xie S, Metcalfe R, Dunn E, Morton C, Huang S, Puhalovich T, Du Y, Wittlin S, Nie S, Luth M, Ma L, Kim M, Pasaje C, Kumpornsin K, Giannangelo C, Houghton F, Churchyard A, Famodimu M, Barry D, Gillett D, Dey S, Kosasih C, Newman W, Niles J, Lee M, Baum J, Ottilie S, Winzeler E, Creek D, Williamson N, Parker M, Brand S, Langston S, Dick L, Griffin M, Gould A, Tilley L, Science 376(6597):1074-1079 (2022) DOI

SASDMD4 – Plasmodium falciparum tyrosyl-tRNA synthetase, S234C mutant

Tyrosine--tRNA ligase (S234C)
MWexperimental 84 kDa
MWexpected 87 kDa
VPorod 124 nm3
log I(s) 2.50×10-2 2.50×10-3 2.50×10-4 2.50×10-5
Tyrosine--tRNA ligase (S234C) small angle scattering data  s, nm-1
ln I(s)
Tyrosine--tRNA ligase (S234C) Guinier plot ln 2.51×10-2 Rg: 3.7 nm 0 (3.7 nm)-2 s2
(sRg)2I(s)/I(0)
Tyrosine--tRNA ligase (S234C) Kratky plot 1.104 0 √3 sRg
p(r)
Tyrosine--tRNA ligase (S234C) pair distance distribution function Rg: 3.8 nm 0 Dmax: 12 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Tyrosine--tRNA ligase (S234C) OTHER model
Synchrotron SAXS data from solutions of P. falciparum tyrosyl-tRNA synthetase, S234C mutant in 20 mM Tris-HCl, 150 mM NaCl, 0.5 mM TCEP, 0.1% sodium azide, pH 7.4 were collected on the SAXS/WAXS beam line at the Australian Synchrotron (Melbourne, Australia) using a Pilatus3 S 2M detector at a sample-detector distance of 2.2 m and at a wavelength of λ = 0.1078 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography co-flow SEC-SAXS was employed. The SEC parameters were as follows: A sample was injected at a 0.40 ml/min flow rate onto a GE Superdex 200 Increase 5/150 column at 20°C. 14 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.

Sample injection volume = UNKNOWN

Tyrosine--tRNA ligase (S234C) (PfYRS S234C)
Mol. type   Protein
Organism   Plasmodium falciparum (isolate 3D7)
Olig. state   Dimer
Mon. MW   43.4 kDa
 
UniProt   Q8IAR7 (2-373)
Sequence   FASTA
 
PDB ID   7ROT