P50 GM073197

JCIMPT NMR Team____________________

Dr. Reto HorstDr. Pedro Serrano

Dr. Pawel Stanczak__________________________________________________

Dr. Kurt Wüthrich

Screening of New DetergentsScreening 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 μL 5 μL

Gilson Liquid Sample Handler

Bruker Automated Sample Changer

1 mm diameter capillary tube vs 5 mm diameter tube

NMR Screening Setup

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

DHPC Fos30

            NMR BUFFER:  20 mM Na Phosphate pH = 6.8100 mM NaCl                  0.3 %    (v/v) NaN3

10 %    (v/v) D2O 2 %   (w/w) DHPC

            NMR BUFFER:  5 mM Na Phosphate pH = 6.810 mM NaCl                  0.3 %    (v/v) NaN3

10 %    (v/v) D2O ≤1 %   (w/w) Detergent

Reconstitution of OmpX from inclusion bodies

Detergent DHPC New Detergents

NMR Sample Preparation of OmpX

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

NMR Sample Preparation of OmpX

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

Monitoring Detergent Concentration Using Micro-coil 1D 1H 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

[15N,1H]-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

[15N,1H]-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

[15N,1H]-TROSY-NMR, cross-sections