;DFHAhnhanca ;avance-version (02/05/31) ;HNCA ;3D sequence with ; inverse correlation for triple resonance using multiple ; inept transfer steps ; ; |F1(HA) -> F2(Ca,t1) -> F2(N,t2) -> F1(HN,t3) ; |F1(HN) -> F2(N,t1) -> F2(Ca,t2) -> F1(HA,t3) ; ; choose F2 NUC : 13C ; choose F3 NUC: 13C ; ;on/off resonance Ca and C=O pulses using shaped pulse ;phase sensitive (t1) ;phase sensitive using Echo/Antiecho gradient selection (t2) ;using constant time in t2 and t1 ;(use parameterset HNCAGP3D) ; PROSOL compatible, but change the following pulses: ; ;make sure Q3 and Q5 are used for Ca and CO (not square pulses) ; ;p24:12 ppm bandwidth REB applied at -1 ppm ;can also use longer REBURP centered at 4 ppm ;sp11:power for HA REBURP ;spnam11: REBURP.1k ; ;paropt p19 to optimize water suppression prosol relations= #include #include #include "d0=3u" "d7=3u" "d11=30m" "d13=4u" "d21=5.54m" ;1/2JNH "d23=12m" ;1/4JCN "d26=2.4m" ;1/4JNH "d24=1.6m" ;1/4JCH ;----1st-NCA-------- "d3=4*d7+3*p14" "d4=d23/2-p14/2-p22/2-p3/2" "TAU3=d3/2+p3/2-p22/2" "d20=d23" "d22=(d20/2-d21-p26-p14/2)" "d25=d20/2-p14/2" "d2=d25-p16-d16" "TAU5=d3/2" ;spectral width calculation: ;put nd0=4 and put the nuc to carbon "in22=in0" "in4=in0" "in25=in0" "in7=1/(4*cnst13*(bf3/1000000))-in0" "d9=p19+d19" ;----2nd-NCA-------- "d5=20u" "d6=(4*d5+3*p14)/2" "d27=d23/2-p14/2-p14/2" "TAU4=(4*d5+3*p14)/2-p22/2-p14/2" "d10=d23/2-p14/2-p22/2-d6" "d29=d23/2-p14/2-d21-4u-p26" "d30=d23/2-p14/2" ;spectral width calculation: ;put nd10=4 and put the nuc to carbon "in29=in10" "in30=in10" "in27=in10" "in5=1/(4*cnst13*(bf3/1000000))-in10" ;"d15=50u" ; REBURP phase compensation for HN "DELTA=d7*2" "DELTA1=p19+d19+4u" "DELTA2=d23-d21-p26" "DELTA3=d26+d24-p24/2-p19-d19-4u" "DELTA4=d26-d24-p4" ;"DELTA5=d21-p24" "DELTA6=d26+d24-p24/2" "DELTA5=d21-d26-d24-p24/2" "DELTA7=d21-d26-d24-p24/2-d9" "DELTA8=d7" "DELTA9=(d26+d24)/2" "DELTA10=(d26-d24)/2" "DELTA11=(d26+d24)/2+p22/2" "DELTA12=(d26-d24)/2-p22/2" "TAU1=(p19+d19)/2" "TAU=p24/2" "TAU2=p16+d16" "spoff2=0" "spoff3=0" "spoff5=bf2*(cnst21/1000000)-o2" "spoff8=0" "spoff11=bf1*(cnst11/1000000)-o1" aqseq 321 1 d11 ze d11 pl16:f3 pl12:f2 2 d11 do:f3 do:f2 3 d1 pl1:f1 pl2:f2 pl3:f3 50u UNBLKGRAD (center (p3 ph1):f2 (p21 ph1):f3) p16:gp10 d16 p1 ph16 d24 pl3:f3 (p4 ph1):f2 DELTA4 (center (p2 ph1) (p22 ph1):f3 ) d26 UNBLKGRAD (p1 ph7):f1 p16:gp1 d16 (p21 ph3):f3 ;start NCA 2HzNy->2NyCz d7 (center (p14:sp5 ph1):f2 (p2 ph1):f1) d7 (p14:sp3 ph1):f2 d7 (center (p14:sp5 ph1):f2 (p2 ph12):f1) d7 (p3 ph3):f2 DELTA6 pl0:f1 pl0:f2 (p24:sp11 ph10):f1 DELTA5 pl19:f1 (p26 ph2):f1 d22 cpds1:f1 ph1 (p14:sp5 ph1):f2 d4 (p22 ph11):f3 TAU3 (p14:sp3 ph11):f2 d25 TAU5 (p14:sp5 ph1):f2 d25 pl2:f2 ;-----NCA-CAN---------------------- (center (p21 ph4):f3 (p3 ph15):f2 ) ;--1st--evolution------- d30 pl0:f2 ;--2nd evolution--- TAU1 (p14:sp5 ph1):f2 d27 TAU1 (p14:sp3 ph1):f2 TAU4 d9 (p22 ph8):f3 d10 d9 (p14:sp5 ph1):f2 d29 d9 4u do:f1 (p26 ph2):f1 ; DELTA7 pl0:f1 (p24:sp11 ph14):f1 4u p19:gp2*EA d19 pl2:f2 pl1:f1 DELTA3 d5 (center (p14:sp5 ph1):f2 (p2 ph1):f1) d5 (p14:sp3 ph1):f2 d5 (center (p14:sp5 ph1):f2 (p2 ph12):f1) d5 p19:gp3*EA d19 (center (p1 ph1) (p3 ph5):f2 (p21 ph5):f3 ) ;start PEP DELTA11 pl3:f3 (p4 ph1):f2 DELTA12 (center (p2 ph1) (p22 ph1):f3 ) DELTA9 (p4 ph1):f2 DELTA10 (center (p1 ph2) (p3 ph6):f2 (p21 ph9):f3 ) DELTA11 (p4 ph1):f2 DELTA12 (center (p2 ph1) (p22 ph1):f3 ) DELTA9 (p4 ph1):f2 DELTA10 (p1 ph12) DELTA1 (p2 ph1);debug p19:gp4 d19 pl16:f3 pl12:f2 4u BLKGRAD go=2 ph31 cpd3:f3 cpds2:f2 d11 do:f3 do:f2 mc #0 to 2 F1PH(rd10 & rd29 & rd30 & rd5 & rd27 & ip3, id7 & id22 &id4 & dd25) F2EA(igrad EA & ip6*2 & ip9*2, id10 & id29 & dd30 &dd27 & id5 & ip15*2 & ip31*2) exit ph1=0 ph2=1 ph21={0}*16 ;{1}*16 ph4 = 0 2 ph3= 0 0 2 2 ph5=0 0 0 0 0 0 0 0 2 2 2 2 2 2 2 2 ph6=1 1 1 1 1 1 1 1 3 3 3 3 3 3 3 3 ph9=3 3 3 3 3 3 3 3 1 1 1 1 1 1 1 1 ph7=3 ph8=0 0 0 0 ph12=2 ph10=0 0 ph14=0 ph11=0 ph15=0 0 0 0 2 2 2 2 ph16={0}*16 ph18={0}*16 ph31=0 2 2 0 2 0 0 2 2 0 0 2 0 2 2 0 ;pl0 : 120dB ;pl1 : f1 channel - power level for pulse (default) ;pl3 : f3 channel - power level for pulse (default) ;pl16: f3 channel - power level for CPD/BB decoupling ;pl12: f2 channel - power level for CPD/BB decoupling ;pl19: f1 channel - power level for CPD/BB decoupling ;sp1: f1 channel - shaped pulse 90 degree (H2O on resonance) ;sp3: f2 channel - shaped pulse 180 degree (Ca on resonance) ;sp5: f2 channel - shaped pulse 180 degree (C=O off resonance) ;p1 : f1 channel - 90 degree high power pulse ;p2 : f1 channel - 180 degree high power pulse ;p11: f1 channel - 90 degree shaped pulse [2 msec] ;p13: f2 channel - 90 degree shaped pulse ;p14: f2 channel - 180 degree shaped pulse ;p16: homospoil/gradient pulse [600u-1 msec] ;p19: encoding/decoding gradient (paropt) [900u] ;p21: f3 channel - 90 degree high power pulse ;p22: f3 channel - 180 degree high power pulse ;p26: f1 channel - 90 degree pulse at pl19 ;d0 : incremented delay (F1 in 3D) [3 usec] ;d1 : relaxation delay; 1-5 * T1 ;d10: incremented delay (F2 in 3D) = d23/2-p14/2 ;d11: delay for disk I/O [30 msec] ;d13: short delay [4 usec] ;d16: delay for homospoil/gradient recovery ;d19: delay for encode/decode gradient recovery ;d21: 1/(2J(NH) [5.5 msec] ;d23: 1/(4J(NCa) [12 msec] ;d26: 1/(4J'(NH) [2.3 msec] ;d29: incremented delay (F2 in 3D) = d23/2-p14/2-p26-d21-4u ;d30: decremented delay (F2 in 3D) = d23/2-p14/2 ;cnst11: HA REBURP offset (-1ppm for 12ppm BWidth) ;cnst13: 15N spectral width ;cnst21: CO chemical shift (offset, in ppm) ;cnst22: Calpha chemical shift (offset, in ppm) ;o2p: Calpha chemical shift (cnst22) ;in0: 1/(8 * SW(N)) = DW(N)=DW(CA)*2 ;nd0: 4 ;in10: 1/(8 * SW(N)) = DW(N)=DW(CA)*2 ;nd10: 4 ;in29: = in10 ;in30: = in10 ;NS: 8 * n ;DS: >= 16 ;td1: number of experiments in F1 ;td2: number of experiments in F2 td2 max = 2 * d30 / in30 ;FnMODE: States-TPPI (or TPPI) in F1 ;FnMODE: echo-antiecho in F2 ;cpds1: decoupling according to sequence defined by cpdprg1 ;cpd3: decoupling according to sequence defined by cpdprg3 ;cpd2: decoupling according to sequence defined by cpdprg2 ;pcpd1: f1 channel - 90 degree pulse for decoupling sequence ;pcpd3: f3 channel - 90 degree pulse for decoupling sequence ;pcpd2: f2 channel - 90 degree pulse for decoupling sequence ;use gradient ratio: gp 2 : gp 3 : gp4 ; -7: -3 : 1 ;for z-only gradients: if p16=600u, p19=900u ;gpz1: 30% ;gpz2: -65.7 ;gpz3: -28.15 ;gpz4: 9.5 ;gpz10: 67 ;use gradient files: ;gpnam1: SINE.100 ;gpnam2: SINE.50 ;gpnam3: SINE.50 ;gpnam4: SINE.50 ;gpnam10: SINE.100 ;from: $Id: hncagp3d,v 1.12 2002/06/12 09:04:45 ber Exp $