;HncocaNH ;modified from hncacogpwg3d ;avance-version (02/05/31) ;HN(COCA)NH ;3D sequence with ; sequential correlation for triple resonance using multiple ; inept transfer steps ; ; F1(Hi+1,t1) -> F3(Ni+1) -> F2(COi) ; -> F2(Cai) -> F3(Ni,t2) -> F1(Hi,t3) ; ;on/off resonance Ca and C=O pulses using shaped pulse ;phase sensitive (t1) ;phase sensitive (t2) ;using semi-constant time in t1 ;using semi-constant time in t2 ;water suppression using watergate sequence ;(use parameterset HNCACOGPWG3D) ; ;(Z-Y.J. Sun, D.P. Frueh, P. Selenko, J.C. Hoch & G. Wagner, J.B. NMR. 33, 43-50 (2005)) ; ;HNCOCANH, strong intra peak with sequential HN connectivity ;optional 2D decoupling ; ;optimal NCA transfer 15.6ms ;Ni+1 refocusing during Ca->N transfer ;optimal d24 = 25ms for Ca seletive exp of protein < 10kD ; ;use d24 = 16ms for best overall experiments ;C13 chemical shifts relative to DSS ;for Ca selective exp (750MHz): p24=1039us cnst22=55ppm ; simulated C13 bandwidth=4148Hz (22ppm) for Q3 shape pulse ;for Ca+ (ZeBra) selective exp (750MHz): p24=737us cnst22=51ppm ; simulated C13 bandwidth=5845Hz (31ppm) for Q3 shape pulse ;for Cab non-selective exp: p24=p14 cnst22=45ppm ; ;l20=1 for 1D test ;l20=2 and set either TD1 or TD2 = 0 for 2D planes ;l20=3 for non-uniformly sampled 3D data set ; ;make sure nd0 = nd10 = 2 ; ;States for H(t1) to shift half spectrum ;so Hi+1 spectrum width can be reduced by half (HN region only) prosol relations= #include #include #include define delay PUL14 "p2=2*p1" "d11=30m" "d21=5.5m" "d22=4.5m" "d23=12m" ;"d24=16m" "d25=13.7m" "d26=2.3m" "d27=15.6m" "d13=d26+p22/2+3u" "d14=3u" "d15=d13-p22-3u" "d28=d25-p14-3u-p14/2" "d29=3u" "d30=3u" "d31=d28-p26-d21-4u" "l3=l1*l2*l20" "in13=d13/l1-0.005u" "in14=in0-in13-(d21/2-d26)/l1" "in15=in0+(d21/2-d26)/l1" "in28=d28/l2-0.005u" "in29=in10-in28-(d27-d25)/l2" "in30=in28+(d27-d25)/l2" "in31=in10-in28" "DELTA1=d23-d21-p26" "DELTA2=d22-p14-4u" "DELTA3=d26-p16-d16-p27*2.385-d18*5+p22/2-4u" "DELTA4=d18-p22/2" "DELTA5=d24-p14-4u" "DELTA6=DELTA5-d22" "CEN_CN1=p14/2-p22/2" "PUL14=p14" "spoff2=0" "spoff3=0" "spoff5=bf2*((cnst21-cnst22)/1000000)" "spoff7=bf2*((cnst22-cnst21)/1000000)" "spoff8=0" "spoff9=0" aqseq 321 1 d11 ze d11 LOCKDEC_ON d11 pl17:f4 2 d11 do:f3 3 d11 pl1:f1 H2_LOCK 6m LOCKH_OFF d1 50u LOCKH_ON 20u H2_PULSE ; CO on resonance 30u fq=cnst21(bf ppm):f2 d11 pl2:f2 pl3:f3 UNBLKGRAD ; purge pulse, destroy 15N, 13C resonance 4u (p21 ph1):f3 4u (p3 ph1):f2 4u p16:gp0 d16 pl0:f2 4u pl0:f1 (p11:sp1 ph13):f1 4u 4u pl1:f1 ; begin H'->N' transfer (d26*2) p1 ph11 d15 (p22 ph1):f3 d14 p2 ph1 d13 (p1 ph12):f1 ; ----- H'zN'z ----- 4u pl0:f1 (p11:sp1 ph12):f1 4u p16:gp1 d16 ; begin N'->CO transfer (d23*2), refocus H' (d21) (p21 ph3):f3 d21 pl19:f1 (p26 ph2):f1 DELTA1 cpds1:f1 ph1 (center (p14:sp3 ph1):f2 (p22 ph1):f3 ) d23 (p21 ph1):f3 ; ----- N'zCOz ----- ; gradient here optional 4u do:f1 (p26 ph7):f1 p16:gp5 ; new gp? d16 ; begin CO->CA transfer (d22) (p13:sp2 ph4):f2 DELTA2 (p14:sp7 ph1):f2 4u (p14:sp3 ph1):f2 DELTA2 (p14:sp7 ph1):f2 4u (p13:sp8 ph2):f2 ; one transfer step only ; ----- N'zCOzCAz ----- 4u 30u fq=cnst22(bf ppm):f2 ; CA on resonance p16:gp5 d16 (p26 ph2):f1 20u cpds1:f1 ph1 20u cpd4:f4 ; begin CA->N transfer (d23*2), refocus N' and CO (d22) (p13:sp2 ph2):f2 d22 (p14:sp5 ph1):f2 4u DELTA6 (center (p24:sp9 ph1):f2 (p22 ph1):f3 ) DELTA5 4u (p14:sp5 ph1):f2 4u (p13:sp8 ph1):f2 ; one transfer and two refocus steps ; ----- CAzNz ----- 4u do:f4 4u do:f1 (p26 ph7):f1 4u p16:gp2 d16 (p26 ph2):f1 20u cpds1:f1 ph1 ; semi-CT N evolution (reverse), refocus CA (d23*2), (p21 ph5):f3 (d28 p14:sp5 ph1 6u PUL14 PUL14 d29 p14:sp3 ph1 d30 p14:sp5 ph1):f2 (d28 PUL14 6u PUL14 CEN_CN1 p22 ph8):f3 d31 4u do:f1 (p26 ph7):f1 d21 (p21 ph1):f3 ; ----- NzHz --- p16:gp3 d16 pl0:f1 (p11:sp1 ph6):f1 4u 4u pl1:f1 ; refocus N, watergate (p1 ph1) 4u p16:gp4 d16 DELTA3 pl18:f1 p27*0.231 ph20 d18*2 p27*0.692 ph20 d18*2 p27*1.462 ph20 DELTA4 (p22 ph1):f3 ;ph2? no difference? DELTA4 p27*1.462 ph22 d18*2 p27*0.692 ph22 d18*2 p27*0.231 ph22 DELTA3 p16:gp4 d16 pl16:f3 4u BLKGRAD go=2 ph31 cpd3:f3 if (l20==1) { d11 do:f3 wr #0 } if (l20==2) { d11 do:f3 mc #0 to 2 F1PH(rd28 & rd29 & rd30 & rd31 & ip11, dd13 & id14 & id15 & rp11) F2PH(ip5, dd28 & id29 & id30 & id31) } if (l20==3) { d11 do:f3 wr #0 if #0 zd ; ----------- non-linear sampling starts ----------------- 20u ip5 lo to 3 times 2 20u ip5*2 20u ip11 20u ip13 lo to 3 times 2 20u ip11*2 20u ip13*2 ; -------------- resetting everything --------------------- 20u rd13 20u rd14 20u rd15 20u rd28 20u rd29 20u rd30 20u rd31 20u rp31 20u rp5 20u rp11 20u rp13 ; ------------- State-TPPI for slow dimension ------------ 95 20u ip31*2 20u ip5*2 20u dd28 20u id29 20u id30 20u id31 lo to 95 times c 20u ivc ; --------- State(-TPPI) for fast dimension ------------ 99 20u ip31*2 ; 20u ip11*2 ;not used for States 20u dd13 20u id14 20u id15 lo to 99 times c 20u ivc lo to 3 times l0 } d11 H2_LOCK d11 LOCKH_OFF d11 LOCKDEC_OFF exit ph1=0 ph2=1 ph3=0 ph4=0 0 2 2 ph5=0 0 0 0 2 2 2 2 ph6=2 ph7=3 ph8=0 0 0 0 0 0 0 0 2 2 2 2 2 2 2 2 ph11=0 ph12=1 3 ph13=2 ph20=0 ph21=1 ph22=2 ph23=3 ph31=0 2 2 0 2 0 0 2 0 2 2 0 2 0 0 2 ;pl0 : 120dB ;pl1 : f1 channel - power level for pulse (default) ;pl2 : f2 channel - power level for pulse (default) ;pl3 : f3 channel - power level for pulse (default) ;pl16: f3 channel - power level for CPD/BB decoupling ;pl18: f1 channel - power level for 3-9-19 watergate pulse ;pl19: f1 channel - power level for CPD/BB decoupling ;sp1: f1 channel - shaped pulse 90 degree (H2O on resonance) ;sp2: f2 channel - shaped pulse 90 degree (on resonance) ;sp3: f2 channel - shaped pulse 180 degree (on resonance) ;sp5: f2 channel - shaped pulse 180 degree (C=O off resonance) ;sp7: f2 channel - shaped pulse 180 degree (Ca off resonance) ;sp8: f2 channel - shaped pulse 90 degree (on resonance) ; for time reversed pulse ;sp9: f2 channel - shaped pulse 180 degree (Ca selective) ; sp9 requires higher selectivity than sp3 ; use Q3.1000 shape file ;p1 : f1 channel - 90 degree high power pulse ;p2 : f1 channel - 180 degree high power pulse ;p11: f1 channel - 90 degree shaped pulse [1 msec] ;p13: f2 channel - 90 degree shaped pulse ;p14: f2 channel - 180 degree shaped pulse ;p16: homospoil/gradient pulse [1 msec] ;p21: f3 channel - 90 degree high power pulse ;p22: f3 channel - 180 degree high power pulse ;p24: f2 channel - 180 degree shaped pulse (sp9) ;p26: f1 channel - 90 degree pulse at pl19 ;p27: f1 channel - 90 degree pulse at pl18, 3-9-19 watergate ;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: decremented semi-CT delay [2.3 msec] ;d14: incremented semi-CT delay [3 usec] ;d15: incremented semi-CT delay [2.3 msec] ;d16: delay for homospoil/gradient recovery ;d18: 72us (750MHz), delay for binomial water suppression ; d18 = (1/(2*d)), d = distance of next null (105us for 500MHz) ;d21: 1/(2J(NH) [5.5 msec] ;d22: 1/(4J(CaCO) [4.5 msec] ;d23: optimal NCO transfter period/2 [12 msec] ;d24: seq. transfer period/2, 16ms default [16 ms] ;d25: optimal CAN transfer period/2 [13.7 ms] ;d26: 1/(4J(NH) [2.3 msec] ;d27: 1/(4J(NCa) [15.6 msec] ;d28: decremented delay (F2 in 3D) [~=d25] ;d29: incremented delay (F2 in 3D) [3u] ;d30: incremented delay (F2 in 3D) [3u] ;d31: incremented delay (F2 in 3D) [~=d25] ;cnst21: CO chemical shift (offset, in ppm) ;cnst22: Calpha chemical shift (see pulseprogram) ;o2p: CO chemical shift (cnst21) ;in0: 1/(2 * SW(H')) = DW(H') nd0=2 ;nd0: 2 ;in10: 1/(2 * SW(N)) = DW(N) nd10=2 ;nd10: 2 ;l0: sampling schedule size (vclist) ;l1: >(d13+d21/2-d26)/in0, max inc td1/2 (vclist) ;l2: >(d28+d27-d25)/in10, max inc td2/2 (vclist) ;l3: dummy variable, not used ;l20: switch 1-1D/2-2D/3-3D exp data sets ;DS: >= 16 ;td1: number of experiments in F1 ;td2: number of experiments in F2 ;FnMODE: States-TPPI (or TPPI) in F1 ;FnMODE: States-TPPI (or TPPI) in F2 ;cpds1: decoupling according to sequence defined by cpdprg1 ;cpd3: decoupling according to sequence defined by cpdprg3 ;pcpd1: f1 channel - 90 degree pulse for decoupling sequence ;pcpd3: f3 channel - 90 degree pulse for decoupling sequence ;use gradient ratio: gp0 : gp1 : gp2 : gp3 : gp4 : gp5 ; 37 : 50 : 40 : 60 : 30 : 53 ;for z-only gradients: ;gpz0: 37% ;gpz1: 50% ;gpz2: 40% ;gpz3: 60% ;gpz4: 30% ;gpz5: 53% ;use gradient files: ;gpnam0: SINE.100 ;gpnam1: SINE.100 ;gpnam2: SINE.100 ;gpnam3: SINE.100 ;gpnam4: SINE.100 ;gpnam5: SINE.100 ;$Id: hncacogpwg3d,v 1.1 2002/06/12 09:04:45 ber Exp $