Legionella pneumophila macrophage infectivity potentiator protein appendage domains modulate protein dynamics and inhibitor binding

Wiedemann C, Whittaker J, Carrillo V, Goretzki B, Dajka M, Tebbe F, Harder J, Krajczy P, Joseph B, Hausch F, Guskov A, Hellmich U, International Journal of Biological Macromolecules :126366 (2023) DOI

SASDSZ6 – Outer membrane protein MIP (LpMIP) with bound inhibitor JK095

Outer membrane protein MIP
MWI(0) 44 kDa
MWexpected 46 kDa
VPorod 65 nm3
log I(s) 8.03×103 8.03×102 8.03×101 8.03×100
Outer membrane protein MIP small angle scattering data  s, nm-1
ln I(s)
Outer membrane protein MIP Guinier plot ln 8.03×103 Rg: 3.1 nm 0 (3.1 nm)-2 s2
(sRg)2I(s)/I(0)
Outer membrane protein MIP Kratky plot 1.104 0 √3 sRg
p(r)
Outer membrane protein MIP pair distance distribution function Rg: 3.1 nm 0 Dmax: 10.2 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Outer membrane protein MIP SREFLEX model
Synchrotron SAXS data from solutions of outer membrane protein MIP (LpMIP) with bound inhibitor JK095 in 20 mM Tris-HCl, 150 mM NaCl, 10 mM DTT, pH 7.5 were collected on the EMBL P12 beam line at PETRA III (DESY; Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 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 45.00 μl sample at 10 mg/ml was injected at a 0.30 ml/min flow rate onto a GE Superdex 200 Increase 5/150 column at 20°C. 2400 successive 0.250 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.

The PubChem entry for the Inhibitor JK095 can be found under: https://pubchem.ncbi.nlm.nih.gov/compound/118257702

Outer membrane protein MIP (LpMIP)
Mol. type   Protein
Organism   Legionella pneumophila subsp. pneumophila (strain Philadelphia 1 / ATCC 33152 / DSM 7513)
Olig. state   Dimer
Mon. MW   22.8 kDa
 
UniProt   Q5ZXE0 (21-233)
Sequence   FASTA
 
PDB ID   1FD9