/* pfg_C_hacaconh_se1.c family of HC(CO)NH straight-through triple resonance experiments Copyright 1995 Protein NMR Laboratory - Center for Advanced Biotechnology and Medicine, Rutgers University C. B. Rios, M. Tashiro, W. Feng, and G. T. Montelione This pulse sequence executes four different triple resonance experiments depending on the settings of the parameters nuc_t1 and nloop: experiment CABM name nuc_t1 nloop H(CCO)NH H-CO-BB H1 0 (H)C(CO)NH C-CO-BB C13 0 H(CCO)NH-TOCSY H-CO-TOCSY H1 >0 C(CCO)NH-TOCSY C-CO-TOCSY C13 >0 These pulse sequences are described in: -Montelione et al, J.Am.Chem.Soc. 1992: 114:10974 - 10975. -Lyons et al., Biochemistry 1993, 32: 7839 - 7845 -Tashiro et al. J. Biomol. NMR 1995, 6: 211-216. -Feng et al. J. Biomol. NMR 1996, 8: 98 - 104. The experiment has been implemented on a Unity 600 spectrometer with 4 channels. If the parameter ch4='y', channel 3 is used for C-alpha frequencies and channel 4 is used for C' frequencies. Carbonyl decoupling is executed with sinx decoupling, but can also be done with SEDUCE or other band-selective decoupling scheme. PLANNED IMPROVEMENTS: NOTE: Dummy statements have been inserted at appropriate points in the code for A) 7.22.97 rn A) The sequence can be modified for 3 channel system by setting ch4='n' and using convolute to define frequency shifted pulses shco90 and shco180 instead, while keeping shca90 and shca180 defined as hard pulses centered on C-alpha. B) Channel 4, if present, can be used for 2H decoupling. C) Small-angle phase-shifts to compensate for Bloch-Seigert effects. ************************************************************************* DEVELOPMENT ISSUES Ported from Unity 500 to Inova 600 7.23.97 rn NOTE: the original versions of this experiment allowed for concat2=n using non-CT N15 freq labeling. We usually run this with concat2=y, giving CT-N15 freq labeling. The current version does not support concat2=n, but some incomplete code is left for this option. for nuc_t1 = 'C13', w1 dimension is C13 for nuc_t1 = 'H1', w1 dimension is H1 includes sinc C' decoupling during Tc when nloop > 0 and nuc_t1 = 'C13' Verification of sensitivity enhancement in C-CO-BB version. Checked s/y in se='y' vs se='n' gtm, ss,cbr 1.3.95 see ss.II.13 sample: [C65A,C72A]-RNaseA Peak se='n' se='y' enhancement 1. 85 9.4 22.7 2.4 2. 74 20.1 30.2 1.5 3. 98 45.8 72.6 1.6 4. 49 38.5 54.6 1.4 If nloop =0, the center of the w1 frequency axis will be set to dof2 = CA frequency. If nloop > 0, The center of w1 frequency axis will be set to dofb = center of aliphatics. You should also use zfilt = 10 to 200 usec (typically 200 usec) to suppress cosy peaks with unreliable signs. The following has not been implemented: NOTE - concat2=n can be used for non-CT 15N freq labeling but must double check sense and all powers for concat2 = n Processing: 2D Spectra - for phase = 1,2 states-tppi in t1. Transform with: wft2d('ni',1,0,0,0,0,0,1,0) in t1 - phase2 = 1,2 nagayama in t2 wft2d('ni2',1,0,1,0,0,-1,0,1) or ('ni2',1,0,0,1,0,1,1,0) in t2 - for phase = 3 tppi in t1. Transform with: wft2d('ni',1,0,1,0,0,0,1,0) in t1 3D Planes - for H-H Planes: wft2d('ni',1,1,0,1,0,0,0,0,0,0,0,0,0,1,0,1,0) INOVA Parameters (revised for 4-channel from 3-channel Unity sequence 6.29.97 rn) Pulses and powers: pw: 90 1H pulse at tpwr on TODEV tpwr: power for 90 1H pulse on TODEV pwCA90l: power for selective 65usec 90 on DO2DEV pwCA180l: power for selective 58usec 180 on DO2DEV pwCO90l: power for selective 90 (CO or D dec?) on DO3DEV pwCO180l: power for selective 180 (CO or D dec?) on DO3DEV pwCA90: 90 pulse at pwCA90l on DO2DEV pwCA180: 180 pulse at pwCA180l on DO2DEV pwCO90: 90 pulse at pwCO90l on DO3DEV pwCO180: 180 pulse at pwCO180l on DO3DEV C90lvl: full power for simultaneous 90 on DO2DEV C90: 90 degree pulse at C90lvl on DO2DEV pwNlvl: power for N on DODEV pwN: 90 degree pulse for N at pwNlvl on DODEV shco90: filename of selective 90 on C' on DO3DEV shca90 filename of selective 90 on CA on DO2DEV shco180: filename of selective 180 on C' on DO3DEV shca180: filename of selective 180 on CA on DO2DEV Decoupling: tpwr_t1: power level for 1H decoupling during t1 on TODEV tpwr_t2: power level for 1H decoupling during t2 on TODEV tseq_t1: filename of multipulse 1H decoupling during t1 on TODEV tseq_t2: filename of multipulse 1H decoupling during t2 on TODEV tres_t1 and tres_t2: resolution of tseq files tmf_t1: tmf degree pulse corresponding to tpwr_t1 on TODEV tmf_t2: tmf corresponding to tpwr_t2 on TODEV dpwr_t1 power level for 15N decoupling during t1 on DODEV dseq_t1: filename of multipulse 15N decoupling during t1 on DODEV dres_t1: resolution of dseq_t1 file dmf_t1: dmf degree pulse corresponding to dpwr_t1 on DODEV dpwr2_t2 power level for Ca (or BB) decoupling during t2 on DO2DEV dseq2_t2: filename of multipulse Ca (or BB) decoupling during t2 on DO2DEV dres2_t2: resolution of dseq2_t2 file dmf2_t2: dmf corresponding to dpwr2_t2 on DO2DEV dpwr3_t1 power level for CO (2H?) decoupling during t1 on DO3DEV dseq3_t1: filename of multipulse CO decoupling during t1 on DO3DEV dres3_t1: resolution of file dmf3_t1: dmf degree pulse corresponding to dpwr3_t1 on DO3DEV Delays, Offsets, Phase Selection, Experiment Selection, Misc: gzlvl1: amplitude of PFG pulse gt1: unit length of PFG pulse quant1: MQ selection parameters for N15 quant1 = 9.9 to 10.1 phase: phase=1,2 for States-TPPI phase=3 for TPPI. phase2: phase2=1,2 selects n- and p-type 15N coherence pathways recover PFG recovery delay zfilt1 z-filter delay zz-filter (usually 0) zfilt2 z-filter delay zz-filter (usually 0) zfilt3 z-filter delay before and after 13C spin lock advisable to use 500 microsec for best phasing H1frq center on H-beta freq (optimize) tof center on HN dof decoffset centered on NH on DODEV dof2 dec2offset C-alpha frequency (on DO2DEV) dof3 dec offset centered on CO (2H??) frequency (on DO3DEV) dofb dec offset aliphatic (C-alpha/C-beta) frequency dofc dec offset (CA + CO) / 2 tauCH: 1/4(J_C-H) ~1.7 ms taufocCH: ~2.3 ms (optimized for CH and CH2) tauCACO: 1/4(J_Calpha-C') 3 to 5 ms tauNH: 1/4(J_N-H) ~2.7 ms tauNCO: 1/4(J_N-C') 10 - 19 ms taue: hardwired to gt1+recover Tc: N/4(J_Calpha-C') 3 to 5 ms for N=1; 13C ct t1-freq labeling Tc Constant time for 13C freq labeling during t1 Tn Constant time for 15N freq labeling during t2 nloop flag to implement ct freq labeling code in t1 nloop=0 gives ct 13C freq labeling and enhanced S/N by concatonating coherence evolution periods use in H-CO-BB, C-CO-BB. nloop>0 gives non-ct 13C freq labeling and must be used in H-CO-TOCSY, C-CO-TOCSY alfa1 corrects lp1, usemacro calfa1 se se = 'y' for sensitivity enhanced se = 'n' without sensitivity enhancement This may be desirable if you have water echoes ch4 ch4 = 'y' for using 4th synthesizer for C' pulses ch4 = 'n' for using SLP for C' pulses NOT IMPLEMENTED: concat2 flag to implement ct freq labeling code in t2 concat2='y' gives ct 15N freq labeling and enhanced S/N by concatonating coherence evolution periods use this as default concat2='n' gives non-ct 15N freq labeling */