P50 GM JCIMPT NMR Team ____________________ Dr. Reto Horst Dr. Pedro Serrano Dr. Pawel Stanczak __________________________________________________ Dr. Kurt Wüthrich

Screening of New Detergents Dr. Reto Horst Dr. Qinhai Zhang Michael Geralt Financial Support through JCIMPT (PI Dr. Ray Stevens) by the NIH Roadmap Initiative and through JCSG (PI Dr. Ian Wilson) by the NIH PSI.

600 μL5 μL Gilson Liquid Sample Handler Bruker Automated Sample Changer 1 mm diameter capillary tube vs 5 mm diameter tube NMR Screening Setup

[ 15 N, 1 H]-TROSY spectra of refolded [ 2 H, 15 N]-labeled OmpW in: DHPCFos30

NMR BUFFER: 20 mM Na Phosphate pH = mM NaCl 0.3 % (v/v) NaN 3 10 % (v/v) D 2 O 2 % (w/w) DHPC NMR BUFFER: 5 mM Na Phosphate pH = mM NaCl 0.3 % (v/v) NaN 3 10 % (v/v) D 2 O ≤1 % (w/w) Detergent Reconstitution of OmpX from inclusion bodies Detergent DHPCNew Detergents

NMR Sample Preparation of OmpX

Micro-scale 1D 1 H NMR Measurement of Detergent Concentration

NMR Sample Preparation of OmpX

Monitoring Detergent Concentration Using Micro-coil 1D 1 H NMR

Impact of Protein Concentration

1.7 % Fos-10 [OmpX] = 0.84 mM 3.3 % Fos-10 [OmpX] = 1.04 mM 28.5 % Fos-10 [OmpX] = 0.77 mM [ 15 N, 1 H]-TROSY-NMR

1.7 % Fos-10 [OmpX] = 0.84 mM 3.3 % Fos-10 [OmpX] = 1.04 mM 28.5 % Fos-10 [OmpX] = 0.77 mM [ 15 N, 1 H]-TROSY-NMR, cross-sections

1.7 % Fos-10 [OmpX] = 0.84 mM 3.3 % Fos-10 [OmpX] = 1.04 mM 28.5 % Fos-10 [OmpX] = 0.77 mM [ 15 N, 1 H]-TROSY-NMR, cross-sections