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P50 GM073197

P50 GM073197. 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

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P50 GM073197

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  1. P50 GM073197 JCIMPT NMR Team ____________________ Dr. Reto Horst Dr. Pedro Serrano Dr. Pawel Stanczak __________________________________________________ Dr. Kurt Wüthrich

  2. 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.

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

  4. [15N, 1H]-TROSY spectra of refolded [2H, 15N]-labeled OmpW in: DHPC Fos30

  5. Reconstitution of OmpX from inclusion bodies Detergent DHPC New Detergents NMR BUFFER: 5 mM Na Phosphate pH = 6.8 10 mM NaCl 0.3 % (v/v) NaN3 10 % (v/v) D2O ≤1 % (w/w) Detergent NMR BUFFER: 20 mM Na Phosphate pH = 6.8 100 mM NaCl 0.3 % (v/v) NaN3 10 % (v/v) D2O 2 % (w/w) DHPC

  6. NMR Sample Preparation of OmpX

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

  8. NMR Sample Preparation of OmpX

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

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

  11. Impact of Protein Concentration

  12. [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

  13. [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

  14. [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

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