;hbacbanhf.fa ;3D/4D HBCB/HACANNH with Watergate (3-9-19) reverse INEPT ; and optional 2H decoupling ;Based on Nietlispach et al., J.Am. Chem. Soc. 1996, 118, 407-415 ;Bruker Avance/Xwin-nmr version ;Written up by F. Abildgaard, NMRFAM (abild@nmrfam.wisc.edu) ; ; $Id: hbacbanhf.fa,v 1.3 1999/05/12 02:36:50 abild Exp abild $ ; ; Disclaimer: This pulse program is provided "as is" for your ; information. Support for the use of this pulse program is only ; provided to users of the National Magnetic Resonance Facility ; at Madison (NMRFAM). Users of this pulse program employ it at ; their own risk. Neither NMRFAM nor University of Wisconsin-Madison ; are liable for any physical or other damage incurred during the ; use of this pulse program. ; ;f1: 1H, f2: 15N, f3: 13C, f4: 2H (channel assignments may be changed below) ;Gradient program: fahbacbanhf ;o1p: 4.7ppm ;o2p: 118ppm ;o3p: use fq3list fahbacbanhf.C (46ppm,57ppm) ;o4p: 4.5ppm ; ;d1: relaxation delay=d1+0.1003s ;p1 90 H at pl1 ;p10 90 H1 SEDUCE-1 shape at power sp1, offset 0, 2ms (if SHAPED defined), ; or p10 90 H1 weak regular pulse at power pl10, 2ms (if SHAPED not defined). ; Check for phase difference between hard H1 and soft H1 ; pulses and set ph10 accordingly. ;p11 90 H at pl11 (H1 composite decoupling) ;p2 90 N at pl2 ;p3 90 Cab at pl3 (for 90 Cab) ;p5 180 CO SEDUCE1 shape at power sp5, offset CO-Cab ;p6 90 Cab at pl6 (for 180 Cab) ;p9 90 Ca at pl9 (for 180 Ca) ;p25 90 2H pulse at pl15 (optional) ;pl0 120dB ;d21: 1/(2*dNu), dNu=(ChSh(NH)-ChSh(H2O))*bf1, 300us at 500MHz ; ;H1 Waltz-16y or DIPSI-2y (cpdprg1), using p90 (PCPD1) at pl11 ;N15 Waltz-16 (cpdprg2), using p90 (PCPD) at pl12 ;C' Compensated SEDUCE1 decoupling of C', using pcpd at sp3, ; shape seduce1c5/6, offset 0, cpdprg3 waltz16sp3. Select a pulse ; length (PCPD) that gives the right offset (CO-CA) and adjust sp3 ; to make PCPD a 90 dgr pulse. ;H2 Waltz-16 (cpdprg5), using p90 (PCPD) at pl15 ;H1 evolution: ; in0, SW(H)=1/(2*in0) ; l8 complex points ;C13 evolution: ; in20, SW(C)=1/(2*in20) ; l6 complex points; max. is (TAUB-p5)/in20+1 ;N15 evolution: ; in10=in30, SW(N)=1/(2*in10) ; l4 complex points; max. is (d10/in10)+1 ; N15 chemical shift axis is reversed: set reverse to true. ;ns=32,64,..., ds=32,64,... ; ;Recommendations for gradients: ;p15=400u ;p16=2.0m ;p17=700u ;cnst21=10% (20G/cm) ;cnst22=-26% (-50G/cm) ;cnst23=21% (40G/cm) ;cnst30=1.8 (adjust) for magic angle gradient (Gx/Gz) or 0 for Gz only. ; ;Define one or more of the following options to tailor this pulse program ; to your specific needs. ; ;#define ONE_D ;uncomment for 1D experiment #define N15_EVOL ;comment out for 2D or 3D w/o N15 evolution ;#define C13_EVOL ;comment out for 2D or 3D w/o C13 evolution #define H1_EVOL ;comment out for 2D or 3D w/o H1 evolution #define H_90180 ;uncomment for an initial delay of dw/2 in t1 (H1) #define C_90180 ;uncomment for an initial delay of dw/2 in t2 (C13) ;#define SHAPED ;uncomment if using a shaped flip-back pulse ;#define H2_DEC ;uncomment to enable H2 decoupling ; ;Define channel assignments: #define H f1 #define N f2 #define C f3 #define D f4 ; ;You shouldn't have to worry about anything beyond this point :-) ; ;sanity checks ; #ifdef ONE_D #undef N15_EVOL #undef C13_EVOL #undef H1_EVOL #endif #ifndef C13_EVOL #undef C_90180 #endif #ifndef H1_EVOL #undef H_90180 #endif ; define delay TAUA1 define delay TAUA2 define delay TAUB define delay TAUC define delay TAUD define delay TAUE define delay TAUW define delay TC define delay TC_TB define delay TN define delay C_H_N define pulse H1_90 define pulse H1_180 define pulse H1_S90 define pulse N15_90 define pulse N15_180 define pulse CA_180 define pulse COB_180 define pulse CAB_90 define pulse CAB_180 "d11=100m" ;disk i/o "d12=10u" ;power switching etc. "d13=5u" ;a short delay "d14=60u" ;ip,id etc. "d16=300u" ;gradient recovery #ifdef H_90180 "d0=in0*0.5+d13" #else "d0=d13" #endif #ifdef C_90180 "d20=in20*0.5+d13" #else "d20=d13" #endif "TAUA1=1.5m-p15-d13*3-p6*2" "TAUA2=1.5m-p15-d13" "TAUB=2.2m" "d3=3.3m" "TC=d3+p6*2-p5-d12*2" "TC_TB=d3-TAUB-d13*5-d12*2-p1*2-p11" "TAUC=11.0m" "TAUD=5.4m" "TAUW=d21" "TAUE=2.2m-TAUW*2.5-H1_90*2.385-p17-d16-d13" "TN=11.2m" "H1_90=p1" "H1_180=p1*2" "H1_S90=p10" "N15_90=p2" "N15_180=p2*2" "CA_180=p9*2" "COB_180=p5" "CAB_90=p3" "CAB_180=p6*2" "C_H_N=N15_90-H1_90" "d22=(TAUW/2)-N15_90" "d10=TN-d13*2-d12" "d30=TN-TAUD-CA_180-p11-d12*2-d13*5" #define H1_DEC_ON d13 \n d12 pl11:H \n p11:H ph0 \n d13 cpds1:H #define H1_DEC_OFF d13 do:H \n p11:H ph2 \n d13 \n d12 pl1:H #ifdef H2_DEC #define H2_DEC_ON d12 pl15:D \n p25:D ph1 \n d13 cpds5:D #define H2_DEC_OFF d13 do:D \n p25:D ph3 #else #define H2_DEC_ON d13 #define H2_DEC_OFF d13 #endif #define SED_ON d13 \n d12 pl0:C \n d13 cpds3:C #define SED_OFF d13 do:C \n d12 pl9:C \n d13 #include #include 1 ze 2 d11 do:N d14 3 d14 d14 d14 d14 4 d14 d14 d14 5 d14 d14 d14 6 d14 d14 d14 7 d14 d14 d14 8 d14 H2_LOCK d14 LOCKH_OFF d1 pl1:H pl2:N ;---------------- set carbon freq. on Cab (46ppm) --------------- d14 fq3:C ;-------------- INEPT to C and H evolution (t1) ----------------- d14 LOCKH_ON d14 H2_PULSE (H1_90 ph21):H d13 GRADIENT15(cnst21) ;400u, 20G/cm TAUA1 pl6:C d0 (CAB_180 ph11):C d0 (H1_180 ph1):H d13 GRADIENT15(cnst21) ;400u, 20G/cm TAUA2 pl3:C (H1_90 ph12):H d12 pl12:N d13 cpds2:N H2_DEC_ON ;------- H1 refocus, CBCA transfer, C evolution (t2) --------------- (CAB_90 ph22):C d13 d12 pl0:C (TAUB) (d20 COB_180:sp5 ph0):C (H1_90 ph12):H d13 (H1_90 ph25):H H1_DEC_ON TC_TB pl6:C (d20 CAB_180 ph13):C (TC) d12 pl0:C (COB_180:sp5 ph0):C d13 do:N d12 pl3:C ;------------------------- INEPT to N -------------------------- (CAB_90 ph0):C d12 pl2:N TAUC pl6:C (CAB_180 ph14 TAUC):C (N15_180 ph0):N d12 pl3:C (CAB_90 ph0):C ;---------------- Switch C frequency to Ca (57ppm) -------------- d13 d14 fq3:C H2_DEC_OFF ;----------- Refocus Ca and constant time evolution on N (t3) ----- SED_ON (N15_90 ph23):N d10 SED_OFF (N15_180 ph15):N d13 (CA_180 ph0):C SED_ON d30 H1_DEC_OFF TAUD (N15_90 ph0):N SED_OFF d13 GRADIENT16(cnst22) ;2m, -50G/cm #ifdef SHAPED d16 pl0:H (H1_S90:sp1 ph10):H ;2ms SEDUCE-1 90 H1 pulse at phase x #else d16 pl10:H (H1_S90 ph10):H ;2ms 90 H1 pulse at phase x #endif ;------------ Reverse INEPT N to amide protons ------------------- d13 d12 pl1:H (H1_90 ph2):H d13 TAUE GRADIENT17(cnst23) ;700u, 40G/cm d16 (H1_90*0.231 ph1):H TAUW (H1_90*0.692 ph1):H TAUW (H1_90*1.462 ph1):H d22 (N15_180 ph0):N d22 (H1_90*1.462 ph3):H TAUW (H1_90*0.692 ph3):H TAUW (H1_90*0.231 ph3):H d13 GRADIENT17(cnst23) ;700u, 40G/cm d16 pl12:N TAUE go=2 ph31 cpds2:N #ifdef ONE_D d11 do:N wr #0 #else d11 do:N wr #0 if #0 zd #endif #ifdef N15_EVOL d14 ip23 lo to 3 times 2 d14 dd10 d14 id30 d14 ip31 d14 ip31 lo to 4 times l4 d14 rd10 d14 rd30 #else d14*7 #endif #ifdef C13_EVOL d14 ip22 lo to 5 times 2 d14 id20 d14 ip31 d14 ip31 lo to 6 times l6 d14 rd20 #else d14*5 #endif #ifdef H1_EVOL d14 ip21 d14 ip25 lo to 7 times 2 d14 id0 d14 ip31 d14 ip31 lo to 8 times l8 #endif d14 H2_LOCK d14 LOCKH_OFF exit ph0=0 ph1=1 ph2=2 ph3=3 ph10=(360) 3 ; phase x (0), adjust for any phase diff. between pl1 and sp1/pl10 ph11=0 2 ph12=1 3 ph13=0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 2 2 2 2 2 2 2 2 3 3 3 3 3 3 3 3 ph14=0 0 2 2 ph15=0 0 0 0 2 2 2 2 ph21=0 ph22=0 ph23=0 0 2 2 ph25=2 ph31=0 2 2 0 0 2 2 0 2 0 0 2 2 0 0 2