Conformational dynamics of the membrane-anchored foldase LipH from Pseudomonas aeruginosa governs recognition and release of its client lipase

Busch M, Loschwitz J, Papadopoulos A, Reiners J Steinchen W, Calvagna V, Smits S, Jaeger K, Kedrov A, Journal of Biological Chemistry :113266 (2026) DOI

SASDYN9 – Lipase chaperone (LipH) VD : Triacylglycerol lipase (LipA) complex

Lipase chaperone
Triacylglycerol lipase
MWexperimental 68 kDa
MWexpected 68 kDa
VPorod 110 nm3
log I(s) 2.05×10-2 2.05×10-3 2.05×10-4 2.05×10-5
Lipase chaperone Triacylglycerol lipase small angle scattering data  s, nm-1
ln I(s)
Lipase chaperone Triacylglycerol lipase Guinier plot ln 2.05×10-2 Rg: 3.1 nm 0 (3.1 nm)-2 s2
(sRg)2I(s)/I(0)
Lipase chaperone Triacylglycerol lipase Kratky plot 1.104 0 √3 sRg
p(r)
Lipase chaperone Triacylglycerol lipase pair distance distribution function Rg: 3.3 nm 0 Dmax: 11.8 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Lipase chaperone (LipH) VD : Triacylglycerol lipase (LipA) complex Rg histogram Rg, nm
Synchrotron SAXS data from solutions of Lipase chaperone (LipH) VD : Triacylglycerol lipase (LipA) complex in 5 mM Tris, 5 mM glycine, 1 mM CaCl2, 5% glycerol,, pH 9 were collected on the EMBL P12 beam line at the PETRA III storage ring (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 8 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.

Lipase chaperone (LipH VD)
Mol. type   Protein
Organism   Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Olig. state   Monomer
Mon. MW   38.3 kDa
 
UniProt   Q01725
Sequence   FASTA
 
Triacylglycerol lipase (LipA)
Mol. type   Protein
Organism   Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Olig. state   Monomer
Mon. MW   30.1 kDa
 
UniProt   P26876
Sequence   FASTA