Kinetic and mechanistic characterization of Mycobacterium tuberculosis glutamyl-tRNA synthetase and determination of its oligomeric structure in solution

Paravisi S, Fumagalli G, Riva M, Morandi P, Morosi R, Konarev P, Petoukhov M, Bernier S, Chênevert R, Svergun D, Curti B, Vanoni M, FEBS Journal 276(5):1398-1417 (2009) DOI

SASDLY5 – Monomer-dimer equilibrium of glutamyl-tRNA synthetase

Glutamate--tRNA ligase

MW^experimental	75	kDa
MW^expected	108	kDa
V^Porod	123	nm³

log I(s) 2.36×10² 2.36×10¹ 2.36×10⁰ 2.36×10^-1

Glutamate--tRNA ligase small angle scattering data

s, nm^-1

Log plot Log-Log plot

ln I(s)

ln 2.36×10² R_g: 3.5 nm 0 (3.5 nm)^-2 s²

(sR_g)²I(s)/I(0)

1.104 0 √3 sR_g

p(r)	R_g: 3.6 nm 0 D_max: 11 nm

Data validation

Experimental data range

p(r) fit to data

Metric

Value

s_min D_max / π

0.6

N_Shannon

Worse

Better

Metric

Value

p_cormap

χ²

0.014
1.344

Worse

Better

Fits and models

log I(s)	s, nm^-1
+3 Δ/σ -3	s, nm^-1

Glutamate--tRNA ligase PDB (PROTEIN DATA BANK) model

Synchrotron SAXS data from solutions of glutamyl-tRNA synthetase in 35 mM HEPES⁄NaOH, pH 6.5 buffer were collected on the EMBL X33 beam line at DORIS III (DESY, Hamburg, Germany) using a MAR 345 Image Plate detector at a sample-detector distance of 2.7 m and at a wavelength of λ = 0.15 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 2.00 mg/ml was measured. Two successive 60 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.

Cell temperature = UNKNOWN. Storage temperature = UNKNOWN

Tags: X33

Glutamate--tRNA ligase (Gts)
Mol. type		Protein
Organism		Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Olig. state		Dimer
Mon. MW		53.9 kDa

UniProt		P9WFV9 (1-490)
Sequence		FASTA

PDB ID		1G59

PDB ID		1GTS