SASDGU5 – The C-terminal cell-surface signaling domain of the Pseudomonas capeferrum anti-sigma regulator PupR in complex with the outer membrane transporter PupB N-terminal signaling domain

MWexperimental 40 kDa MWexpected 32 kDa VPorod 56 nm3 log I(s) 4.42×101 4.42×100 4.42×10-1 4.42×10-2  s, nm-1 Log plot Log-Log plot

ln I(s) ln 4.43×101 Rg: 2.5 nm 0 (2.5 nm)-2 s2 (sRg)2I(s)/I(0) 1.104 0 √3 sRg p(r) Rg: 2.6 nm 0 Dmax: 8.7 nm

Fits and models

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log I(s)  s, nm-1 +3 Δ/σ -3  s, nm-1 Fit validation Metric Value pcormap χ² 0.525 1.282 Worse Better

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log I(s)  s, nm-1 +3 Δ/σ -3  s, nm-1 Fit validation Metric Value pcormap χ² 0 3.612 Worse Better

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Synchrotron SAXS data from solutions of PupR in complex with PupB in 25 mM HEPES, 400 mM LiCl, 10% v/v glycerol, pH 7.5 were collected on the BioCAT 18ID beam line at the Advanced Photon Source (APS), Argonne National Laboratory (Lemont, IL, USA) using a Pilatus3 X 1M detector at a sample-detector distance of 3.5 m and at a wavelength of λ = 0.103 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 500.00 μl sample at 12.8 mg/ml was injected at a 0.60 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 25°C. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of an appropriate solvent-blank was subtracted from the sample SEC-elution peak. Storage temperature = UNKNOWN. X-ray Exposure time = UNKNOWN. Number of frames = UNKNOWN

PupR protein (PupR) Mol. type   Protein Organism   Pseudomonas putida Olig. state   Monomer Mon. MW   24.1 kDa   UniProt   Q52209 (110-324) Sequence   FASTA   Ferric-pseudobactin BN7/BN8 receptor (PupB) Mol. type   Protein Organism   Pseudomonas putida Olig. state   Monomer Mon. MW   8.1 kDa   UniProt   P38047 Sequence   FASTA