;hNcaNH ;modified from hncacogpwg3d and hncocanh ;avance-version (02/05/31) ;(H)N(CA)NH ;3D sequence with ; sequential correlation for triple resonance using multiple ; inept transfer steps ; ; F1(Hi+-1) -> F3(Ni+-1,t1) -> ; -> F2(Cai) -> F3(Ni,t2) -> F1(Hi,t3) ; ;on/off resonance Ca and C=O pulses using shaped pulse ;phase sensitive PH (t1) (TROSY IPAP) ;phase sensitive EA (t2) ;using semi constant time in t1 ;using semi constant time in t2 ; ;R.T. Clubb, V. Thanabal & G. Wagner, J. Magn. Reson. 97, 213-217 (1992) ;(L.E. Kay, G.Y. Xu & T. Yamazaki, J. Magn. Reson. A109, 129-133 (1994)) ;Frueh, D., Sun, J., Vosburg, DA, Hoch, J., Walsh, CT., Wagner, G JACS 2006 ;modified by Jim Sun, 01/05 ;add N' CT evolution ;add N' refocusing during Ca->N transfer ;optimize semi-CT final transfer time ;add SE then TROSY in N period D Frueh + J Sun ;modified for hNNH ;remove 1H decoupling, add EA-TPPI and concatenation ;add water flipback on 750 ;NUS implementation, add ph5 cycling ;add IPAP trosy in N' period, ipap D Frueh + J Sun ;******************************************* ;* * ;* Prosol compatible, but: * ;* use CA selective pulse for p24:sp9 * ;*-----------------------------------------* ;*- non uniform sampling: see general NUS * ;* file on wagner web-site * ;*- use sampsched2d (rnmrtk) to generate * ;* schedule; then sched2vc * ;*- optimze d23, d24 and d25 (see paper) * ;* very rough estimation CA T2 by * ;* uncommenting ;(p24:sp9 ph20:r):f2 and commenting * ;* the line (center (p24:sp9 ph20:r):f2 (p22 ph1):f3 ) * ;* (see comment inside the pulprog) * ;* ;*- note: signal not seen in 1D when auto * ;* correlation peak is present (depends on * ;* the value of d24) * ;* -sensitivity is about half TROSY-HNCACB * ;* -depending on type of board, need to * ;* change NUS (TCU error for high-resolution* ;*low resolution can be achieved by changing * ;* values of l15 and l16 manualy (need to * ;*comment current definitions * ;* * ;* tested on 750, 600, and 900 Bruker AVANCE* ;* enjoy * ;* DF * ;********************************************** prosol relations= #include #include #include define delay PUL14 "p2=2*p1" "d11=30m" "d21=5.5m" ;"d23=12m" ;"d24=d23" ;"d25=d23" "d26=2.3m" "d9=16m" ;SCT delay "d17=d21/2-p1-p11-12u" "d12=d23-p14-3u-p14/2" "d13=3u" "d14=3u" "d15=d12-d21/2-p1-p11-12u" "d28=d25-p14-3u-p14/2-d26" ;concatenation "d29=3u" "d30=3u" "d31=d28-d21/2-p2-p1" "l15=td1" "l16=td2" if (l15 > 1) { "l1=td1/2" } if (l16 > 1) { "l2=td2/2" } "l3=l1*l2" "in12=d12/l1" ;decremented "in13=in0-d9/l1" "in14=d9/l1" "in15=in0-d12/l1" "in28=d28/l2" "in29=in10-d9/l2" "in30=d9/l2" "in31=in10-d28/l2" "DELTA1=d23-d26-p2" "DELTA2=d24-p24/2" "DELTA5=d21/2-p16-d16" "DELTA6=p16+d16+4u+4u-p21/2" "DELTA7=d26-p21/2" "DELTA8=d26-p11-4u-4u" "CEN_CN1=p14/2-p22/2" "CEN_HC1=p14/2-p2/2" "PUL14=p14" "spoff2=0" "spoff3=0" "spoff5=bf2*((cnst21-cnst22)/1000000)" "spoff8=0" "spoff9=0" aqseq 321 1 d11 ze d11 LOCKDEC_ON d11 pl0:f2 pl16:f3 pl17:f4 2 d11 do:f3 3 d11 pl1:f1 H2_LOCK 6m LOCKH_OFF d1 pl1:f1 fq=cnst22(bf ppm):f2 ; CA on resonance d11 pl2:f2 pl3:f3 UNBLKGRAD 20u H2_PULSE ; purge pulse, destroy 15N, 13C resonance 4u ; (p21 ph1):f3 4u (p3 ph1):f2 4u p16:gp1 ; new gp? d16 pl0:f2 4u pl0:f1 ; (p11:sp1 ph22:r):f1 4u 4u pl1:f1 ; begin H'->N' transfer (d26*2) p1 ph1 d26 ;pl3:f3 (center (p2 ph1) (p22 ph1):f3 ) d26 (p1 ph2):f1 ; ----- H'zN'z ----- 4u pl0:f1 (p11:sp1 ph24:r):f1 4u p16:gp1 d16 ; begin N'->CA double transfer, refocus H' decouple CO (p21 ph3):f3 d17 ;1/4J 4u pl0:f1 (p11:sp1 ph22:r):f1 4u 4u pl1:f1 (p1 ph1) (p1 ph29) 4u pl0:f1 (p11:sp1 ph28:r):f1 4u 4u pl1:f1 d15 (p14:sp5 ph1 d14 p14:sp3 ph1 d13 PUL14 PUL14 6u p14:sp5 ph1 d12):f2 (PUL14 d14 PUL14 d13 CEN_CN1 p22 ph8):f3 (p21 ph1):f3 ; ----- N'zCA'zCAz ----- p16:gp5 d16 pl17:f4 ;-------------TMP------------- 20u cpds4:f4 ph1 ; begin CA->N transfer, refocus N' decouple CO (p13:sp2 ph4):f2 DELTA2 ;(p24:sp9 ph20:r):f2 ;to estimate relaxation (center (p24:sp9 ph20:r):f2 (p22 ph1):f3 ) DELTA2 (p13:sp8 ph1):f2 ; ----- CAzNz ----- ;-------------TMP------------- 4u pl1:f1 4u do:f4 4u p16:gp2 d16 ; 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 pl1:f1 p2 ph1 DELTA5 p16:gp12*EA d16 pl1:f1 (p1 ph17):f1 4u pl0:f1 (p11:sp1 ph19:r):f1 4u DELTA8 pl1:f1 (center (p2 ph1) (p14:sp3 ph1):f2 (p22 ph1):f3 ) DELTA8 pl0:f1 (p11:sp1 ph10:r):f1 4u 4u pl1:f1 (center (p1 ph1) (p21 ph2):f3 ) d26 (center (p2 ph1) (p22 ph1):f3 ) DELTA7 (p21 ph18):f3 DELTA6 (p2 ph1) 4u p16:gp13 d16 ;pl16:f3 4u BLKGRAMP go=2 ph31 ;cpd3:f3 if (l20 ==2) { d11 do:f3 mc #0 to 2 F1PH(rd28 & rd29 & rd30 & rd31 & ip3, dd12 & id13 & id14 & id15) F2EA(ip17*2 & ip18*2 & ip19*2 & igrad EA, dd28 & id29 & id30 & id31 & ip5*2 & ip31*2) } if (l20==3) { d11 do:f3 wr #0 if #0 zd ; ----------- non-linear sampling starts ----------------- 1m ip17*2 ;on 750 MHz, 20u works 1m ip18*2 1m ip19*2 1m igrad EA lo to 3 times 2 1m ip3 lo to 3 times 2 1m ip3*2 ; -------------- resetting everything --------------------- 1m rd12 1m rd13 1m rd14 1m rd15 1m rd28 1m rd29 1m rd30 1m rd31 1m rp31 1m rp5 1m rp3 ; ------------- State-TPPI for slow dimension ------------ 95 1m ip31*2 1m ip5*2 1m dd28 1m id29 1m id30 1m id31 lo to 95 times c 1m ivc ; ------------- State-TPPI for fast dimension ------------ 99 1m ip31*2 1m ip3*2 1m dd12 1m id13 1m id14 1m id15 lo to 99 times c 1m ivc lo to 3 times l0 } if (l20 ==1) { d11 do:f3 wr #0 } d11 H2_LOCK d11 LOCKH_OFF d11 LOCKDEC_OFF exit ph1=0 ph2=1 ph3=0 1 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 ph10=0 ph11=0 ph15=0 0 ;2 2 ph16=3 3 ;1 1 ph17=1 ph18=2 ph19=1 ph20=0 ph21=1 ph22=2 ph23=3 ph24=0 ;2 ph28=2 0 ph29=0 2 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 on resonance) ; sp9 requires higher selectivity than sp3 ;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: delay for binomial water suppression ; d18 = (1/(2*d)), d = distance of next null (476 Hz?) ;d21: 1/(2J(NH) [5.5 msec] ;d22: 1/(4J(CaCO) [4 msec] ;d23: 1/(4J(NCa) first period [12 msec] ;d25: 1/(4J(NCa) last period [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 ;cnst21: CO chemical shift (offset, in ppm) ;cnst22: Calpha chemical shift (offset, in ppm) ;o2p: CO chemical shift (cnst21) ;in0: 1/(2 * SW(N')) = DW(N') ;nd0: 2 ;l1: l1 = td1 / 2 ;in10: 1/(2 * SW(N)) = DW(N) ;nd10: 2 ;l2: l2 = td2 / 2 ;in28: =~ d23 / l2 ;in29: =~ in10 ;in30: =~ d23 / l2 ;in31: =~ 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: 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: gp 1 : gp 2 : gp 3 : gp 4 ; 50 : 40 : 60 : 30 ;for z-only gradients: ;gpz1: 50% ;gpz2: 40% ;gpz3: 60% ;gpz12: 80% ;gpz13: 8% ;gpz4: 30% ;use gradient files: ;gpnam1: SINE.100 ;gpnam2: SINE.100 ;gpnam3: SINE.100 ;gpnam4: SINE.100 ;$Id: hncacogpwg3d,v 1.1 2002/06/12 09:04:45 ber Exp $