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