p50 gm073197
DESCRIPTION
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 - PowerPoint PPT PresentationTRANSCRIPT
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