;hncacbf.fa ;3D HNCACB, gradient enhanced with Watergate reverse INEPT (3-9-19), ; minimal H2O saturation and optional 2H decoupling ;Yamazaki et al., J. Am. Chem. Soc. 1994, 116, 11655-11666 ;Wittekind and Mueller, J. Magn. Reson. B 101 201-205 (1993) ;Mori et al., J. Magn. Reson. B 108, 94-98 (1995) ;Bruker Avance/Xwin-nmr version ;Written up by F. Abildgaard, NMRFAM (abild@nmrfam.wisc.edu) ; ; $Id: hncacbf.fa,v 1.2 1999/04/22 19:55:54 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: fahncacbf ;o1p: 4.7ppm, ;o2p: 118ppm, ;o3p: 43ppm, ;o4p:: 4.5ppm ; ;d1: relaxation delay=d1+0.1003s ;p1 90 H1 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 ph21 accordingly. ;p11 90 H1 at pl11 (H1 composite decoupling) ;p2 90 N at pl2 ;p3 90 Cab at pl3 (for 90 Cab), typ. 17kHz at 500MHz ;p25 90 2H pulse at pl15 ;d3: set d3 to 3.5ms for Ca and Cb, or 6.8ms for Cb only. ;pl0 120dB ;d21: 1/(2*dNu), dNu=Nu(NH)-Nu(H2O) ; ;H1 Waltz-16x or DIPSI-2x (cpdprg1), using p90 (PCPD) at pl11 ;N15 Waltz-16 (cpdprg2), using p90 (PCPD) at pl12 ;C' Compensated SEDUCE1 decoupling of C', using p90 (PCPD) at sp3, ; shape seduce1c5/6, offset 0, cpdprg3 waltz16sp3. Select a pulse ; length (PCPD) that gives the right offset and adjust sp3 to make ; PCPD a 90 dgr pulse. ;H2 Waltz-16 (cpdprg5), using p90 (PCPD5) at pl15 ;N15 evolution: ; in10=in30, SW(N)=1/2*in10 ; l4 complex points; max. is (d10/in10)+1 ; Process as States (although data is acquired as States-TPPI) ; N15 chemical shift axis is reversed: set reverse to true. ;C13 evolution: ; in0, SW(C)=1/2*in0 ; l6 complex points ; set cnst0 to 0 (preferably) or 1 to make d0 the smallest possible ; positive delay. cnst0=0 gives (90,-180) phase distortion in F1. ; cnst0=1 gives (270,-540) phase distortion in F1 (use LP to correct). ;ns=8, 16, ..., ds=8, 16,... ; ;Recommendations for gradients: ;p15=500u ;p16=1.0m ;p17=700u ;p18=1.0m ;p19=600u ;p20=100u ;cnst21=6% (4G/cm z) ;cnst22=15% (10G/cm z) ;cnst23=20% (40G/cm combined x,z) ;cnst24=-8% (-5G/cm z) ;cnst25=15% (10G/cm z) ;cnst26=15% (10G/cm z) ;cnst27=12% (8G/cm z) ;cnst28=29% (20G/cm z) ;cnst29=44% (30G/cm z) ;cnst30=1.800 (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 w/o N15 evolution #define C13_EVOL ; comment out for 2D w/o C13 evolution ;#define SHAPED ; uncomment if shaped flip-back pulse ;#define H2_DEC ; uncomment to enable H2 decoupling ;#define SIDECHN ; uncomment for delays optimized for side-chain amide ; ; correlations #define EXPTCORR ; uncomment if you want "expt" to report ; ; the correct expt time (works with XWIN-NMR 2.x) ; ;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 #endif ; aqseq 321 ; define delay TAUA1 define delay TAUA2 define delay TAUB define delay TAUC define delay TAUC1 define delay TAUF1 define delay TAUF2 define delay TAUW 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 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 "d17=50u" ;short gradient recovery "H1_90=p1" "H1_180=p1*2" "H1_S90=p10" "N15_90=p2" "N15_180=p2*2" "CAB_90=p3" "CAB_180=p3*2" "TAUW=d21" "TAUA1=2.25m-p15-d16-d13" "TAUA2=2.25m-TAUW*2.5-H1_90*2.385-p17-d16-d13" #ifdef SIDECHN "TAUB=2.7m" "TN=20.0m" #else "TAUB=5.4m" "TN=12.0m" #endif "TAUC=TN-CAB_180" "TAUC1=TAUC+CAB_180-TAUB-p11-d13-d12" #ifdef H2_DEC "TAUF1=d3-p20-d17-p11-p25-d12*3-d13*7" "TAUF2=TAUF1-d12-d13" #else "TAUF1=d3-p20-d17-p11-d12*3-d13*7" "TAUF2=TAUF1-d13" #endif "d10=TN-d13*2-d12" "d30=TN-TAUB-CAB_180-p11-d12*2-d13*5" #ifdef C13_EVOL "d0=((cnst0*2+1)*in0-CAB_90*1.273-N15_180-d12*2-d13*4)/2" #endif "d22=(TAUW/2)-N15_90" "C_H_N=N15_90-H1_90" #ifdef EXPTCORR "d31=2*TAUA1+2*TAUB+TAUC1+TAUC+2*TAUF1+2*TAUF2+2*TAUA2+4*TAUW" #endif #define H1_DEC_ON d13 \n d12 pl11:H \n p11:H ph1 \n d13 cpds1:H #define H1_DEC_OFF d13 do:H \n p11:H ph3 \n d13 \n d12 pl1:H #define H1_DEC_OFF_Y d13 do:H \n p11:H ph1 \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 pl3: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 d14 6 d14 H2_LOCK d14 LOCKH_OFF #ifdef EXPTCORR #include #endif d1 pl1:H pl2:N d14 LOCKH_ON d14 H2_PULSE (N15_90 ph0):N d13 GRADIENT17(cnst26) ;400u, 10G/cm d16 (H1_90 ph0):H d13 GRADIENT15(cnst21) ;500u, 4G/cm d16 TAUA1 (C_H_N H1_180 ph0):H (N15_180 ph0):N TAUA1 d13 GRADIENT15(cnst21) ;500u, 4G/cm d16 (H1_90 ph1):H d13 #ifdef SHAPED d12 pl0:H (H1_S90:sp1 ph21):H ;2ms SEDUCE-1 90 H1 pulse at phase x #else d12 pl10:H (H1_S90 ph21):H ;2ms 90 H1 pulse at phase x #endif d13 GRADIENT16(cnst22) ;1.0m, 10G/cm d16 SED_ON (N15_90 ph11):N TAUB H1_DEC_ON TAUC1 SED_OFF (N15_180 ph0):N d13 (CAB_180 ph0):C SED_ON TAUC (N15_90 ph0):N H1_DEC_OFF SED_OFF d13 GRADIENT18(cnst24) ;1.0m, -5G/cm d16 H2_DEC_ON H1_DEC_ON (CAB_90 ph22):C SED_ON TAUF1 H2_DEC_OFF H1_DEC_OFF SED_OFF GRADIENT20(cnst28) ;100u, 20G/cm d17 (CAB_180 ph0):C d13 GRADIENT20(cnst28) ;100u, 20G/cm d17 SED_ON H1_DEC_ON H2_DEC_ON TAUF2 SED_OFF (CAB_90 ph1):C #ifdef C13_EVOL SED_ON d0 (N15_180 ph0):N d0 SED_OFF #else d13 #endif (CAB_90 ph23):C SED_ON TAUF1 H2_DEC_OFF H1_DEC_OFF SED_OFF GRADIENT20(cnst29) ;100u, 30G/cm d17 (CAB_180 ph0):C d13 GRADIENT20(cnst29) ;100u, 30G/cm d17 SED_ON H1_DEC_ON H2_DEC_ON TAUF2 SED_OFF (CAB_90 ph24):C H2_DEC_OFF H1_DEC_OFF GRADIENT19(cnst25) ;600u, 10G/cm d16 SED_ON H1_DEC_ON (N15_90 ph13):N d10 SED_OFF (N15_180 ph14):N d13 (CAB_180 ph0):C SED_ON d30 H1_DEC_OFF TAUB (N15_90 ph0):N SED_OFF d13 GRADIENT15(cnst27) ;500u, 8G/cm #ifdef SHAPED d16 pl0:H (H1_S90:sp1 ph21):H ;2ms SEDUCE-1 90 H1 pulse at phase x #else d16 pl10:H (H1_S90 ph21):H ;2ms 90 H1 pulse at phase x #endif d13 d12 pl1:H (H1_90 ph2):H d13 TAUA2 pl1:H 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 TAUA2 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 ip13 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 ip23 d14 ip24 lo to 5 times 2 d14 id0 d14 ip31 d14 ip31 lo to 6 times l6 #endif d14 H2_LOCK d14 LOCKH_OFF exit ph0=0 ph1=1 ph2=2 ph3=3 ph11=0 2 ph13=0 ph14=0 0 0 0 2 2 2 2 ph22=0 0 2 2 ph23=1 1 1 1 3 3 3 3 ph24=0 0 0 0 0 0 0 0 2 2 2 2 2 2 2 2 ph21=(360) 358 ; phase x, adjust for any phase diff. between pl1 and sp1/pl110 ph31=0 2 2 0 0 2 2 0 2 0 0 2 2 0 0 2