The Chlamydia pneumoniae effector SemD exploits its host’s endocytic machinery by structural and functional mimicry

Kocher F, Applegate V, Reiners J Port A, Spona D, Hänsch S, Mirzaiebadizi A, Ahmadian M, Smits S, Hegemann J, Mölleken K, Nature Communications 15(1) (2024) DOI

SASDTT5 – SemDΔAPH bound to Sorting nexin-9, N-terminus - SNX9-SH3

Uncharacterized protein
Sorting nexin-9
MWexperimental 53 kDa
MWexpected 54 kDa
VPorod 94 nm3
log I(s) 7.33×10-3 7.33×10-4 7.33×10-5 7.33×10-6
Uncharacterized protein Sorting nexin-9 small angle scattering data  s, nm-1
ln I(s)
Uncharacterized protein Sorting nexin-9 Guinier plot ln 7.34×10-3 Rg: 3.4 nm 0 (3.4 nm)-2 s2
(sRg)2I(s)/I(0)
Uncharacterized protein Sorting nexin-9 Kratky plot 1.104 0 √3 sRg
p(r)
Uncharacterized protein Sorting nexin-9 pair distance distribution function Rg: 3.5 nm 0 Dmax: 12.2 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Uncharacterized protein Sorting nexin-9 CORAL model
Synchrotron SAXS data from solutions of SemDΔAPH bound to SNX9-SH3 in 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 3% glycerol, pH 8.5 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 100.00 μl sample at 3.3 mg/ml was injected at a 0.60 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 20°C. 2400 successive 0.995 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.

Uncharacterized protein (SemDΔAPH)
Mol. type   Protein
Organism   Chlamydia pneumoniae
Olig. state   Monomer
Mon. MW   35.1 kDa
 
UniProt   Q9Z7M7 (67-382)
Sequence   FASTA
 
Sorting nexin-9 (SNX9-SH3)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   18.8 kDa
 
UniProt   Q9Y5X1 (1-160)
Sequence   FASTA