NMR solution structure of a highly stable de novo heterodimeric coiled-coil
The peptide was chemically synthesized using t-butyloxycarbonyl solid-phase technique
back calculated data agree with experimental NOESY spectrum
SOFTWARE USED WAS CYANA AND ITS INTEGRATED AUTO-ASSIGNMENT MODULE CANDID. THE CHEMICAL SHIFT IDENTIFICATION TOLERANCE AND TRANSPOSE ERRORS WERE SET TO 0.015 PPM IN BOTH PROTON DIMENSIONS. STARTING WITH DISTANCE RESTRAINTS DERIVED FROM MANUALLY ASSIGNED NOE CROSS-PEAKS IN NMRVIEW, A TOTAL NUMBER OF 100 STRUCTURES WERE GENERATED PER CANDID ROUND WITH 8000 STEPS IN THE CYANA ANNEALING PROTOCOL. FOR EACH CANDID RUN, TALOS DERIVED ANGLE RESTRAINTS WERE INCORPORATED (WITH A MINIMUM ERROR SET MANUALLY TO 20 DEGREES) . THE BEST 20 LOW TARGET FUNCTION VALUE STRUCTURES OF CANDIDS FINAL ROUND WERE KEPT AS AN ENSEMBLE REPRESENTATION OF THE FAMILY OF GENERATED STRUCTURES.
DISTANCE GEOMETRY, SIMULATED ANNEALING, CHEMICAL-SHIFT AUTO -ASSIGNMENT, DISTANCE GEOMETRY, SIMULATED ANNEALING
1MM IAAL-E3/K3 COILED-COIL, NA-15N, PH 6.7, 100MM KCL, 50MM KPO4,90%/10% H2O/D2O, 23.1% TFE-D3