Structural insights into the substrate-binding proteins Mce1A and Mce4A from Mycobacterium tuberculosis

Asthana P, Singh D, Pedersen J, Hynönen M, Sulu R, Murthy A, Laitaoja M, Jänis J, Riley L, Venkatesan R, IUCrJ 8(5) (2021) DOI

SASDJV9 – Mammalian cell entry protein 4A (Mce4A39-140)

Mce-family protein Mce4A
MWI(0) 14 kDa
MWexpected 15 kDa
VPorod 22 nm3
log I(s) 9.22×10-3 9.22×10-4 9.22×10-5 9.22×10-6
Mce-family protein Mce4A small angle scattering data  s, nm-1
ln I(s)
Mce-family protein Mce4A Guinier plot ln 9.22×10-3 Rg: 2.1 nm 0 (2.1 nm)-2 s2
(sRg)2I(s)/I(0)
Mce-family protein Mce4A Kratky plot 1.104 0 3 sRg
p(r)
Mce-family protein Mce4A pair distance distribution function Rg: 2.1 nm 0 Dmax: 7.8 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Mce-family protein Mce4A DAMMIN model

log I(s)
 s, nm-1
Mce-family protein Mce4A PDB (PROTEIN DATA BANK) model

log I(s)
 s, nm-1
Mce-family protein Mce4A PDB (PROTEIN DATA BANK) model

Synchrotron SAXS data from solutions of Mammalian cell entry protein 4A (Mce4A39-140) in 50mM MOPS, 350mM NaCl, 10% Glycerol, pH 7 were collected on the B21 beam line at the Diamond Light Source storage ring (Didcot, UK) using a Pilatus 2M detector at a wavelength of λ = 0.1 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 1.00 mg/ml was measured. 28 successive 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.

Synchrotron SAXS data of the Mce domain of Mammalian cell entry protein (Mce) 4A collected at DLS beamline B21 using batch mode method. The molecular mass was determined using Size exclusion chromatography-multi angle light scattering (SEC-MALS) coupled to refractive index measurements and also with the SAXS data. The protein models used were X-ray crystal structure, with missing residues built using Robetta software. The models were fitted against the SAXS data using the in-house program.

Mce-family protein Mce4A (Mce4A)
Mol. type   Protein
Organism   Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Olig. state   Monomer
Mon. MW   15.1 kDa
 
UniProt   I6YC99 (39-140)
Sequence   FASTA