;hnngp3d ;avance-version ;3D HNN (800 MHz) ;o2p = 56.0 ppm ;Panchal et el J Biomol NMR 20, 135, 2001 ;Chugh J et al J Biomol NMR 40, 145, 2008 ;1) exclude Rance-Kay trick ;2) use maximum resolution for t2 including also period k ;3) water flip back and final watergate for water suppression ;sequence checkd and corrected by DMO ;edited by jc and dkg at 800 MHz ;############# HNN suite of experiments ################# ;p34 pulse makes this experiment selective for glycines/alanines/ser-threonines ;p34 = 200us for glycines ;p34 = optimize around 325 us for alanines ;p34 = optimize around 650 us for ser-threonines with SPOFF20 = 1600 Hz (carrier for this pulse shifted to 64 ppm) ;######################################################## ;$OWNER=guest prosol relations = #include #include ;############### constant values ############## "d2=2.25m" ;"d4=2.7m-p2-3u-4u-p16-d16" "d5=2.7m-p11-3u-4u-p16-d16" ;added DMO "d6=5.4m" ;k ;"d7=14m" ;Tn "d11=30m" "d14=p14" ;"d15=12.5m" ;Tcn=12-17m "d16=100u" ;############### center pulses ################ "d3=p21-p1" "d8=(p14-p21*2)*0.5" ;############### incremented time t1 ########## "d0=3u" ;t1/2 "d9=d0+d14+d7+d8" ;t1/2 NOTE that d9<=d8+3u ;############### incremented time t2 ########## "d20=d9-d8" ;t2/2 decreased d20 ;d7=d9= 15ms = Tn "d10=3u" ;t2/2 increased "d21=d10+d9+d8+d7-d6-p1-p21*2-24u" "in9=in0" "in20=in10" "spoff2=0" "spoff3=0" "spoff4=0" "spoff5=bf2*(cnst21/1000000)-o2" ;"spoff20=bf2*(cnst24/1000000)-o2" ;cnst24=64 aqseq 312 1 ze d11 pl13:f3 2 d11 pl13:f3 d1 pl1:f1 20u pl2:f2 50u UNBLKGRAD (p1 ph0) d2 pl3:f3 (d3 p1*2 ph0) (p21*2 ph0):f3 d2 (p1 ph9):f1 3u 20u pl11:f1 (p11:sp1 ph7:r):f1 3u p16:gp1 d16 pl1:f1 (p21 ph1):f3 ;#### Start t1 d0=A ; d7=15m=B ; d9=C=15m-3u ; d6=5.4m=k ######### (d6 p1 ph9 20u pl19 4u cpds1):f1 (d0 p14:sp5 ph0 d7 p14:sp3 ph0):f2 (d0 d14 d7 d8 p21*2 ph0 d9):f3 ;########### end t1 ############ (p21 ph2):f3 4u do:f1 20u pl1:f1 (p1 ph10):f1 4u p16:gp2 d16 (p1 ph9):f1 3u 20u pl19:f1 4u cpds1:f1 (p13:sp2 ph2):f2 d15 ;Tcn = 12-16 ms (d8 p21*2 ph0):f3 (p34:sp20 ph0):f2 d15 ;Tcn = 12-16 ms (p13:sp4 ph0):f2 4u do:f1 20u pl1:f1 (p1 ph10):f1 4u p16:gp3 d16 (p1 ph9):f1 3u 20u pl19:f1 4u cpds1:f1 (p21 ph8):f3 5u ;#### Start t2 d10=F ; d20=D=15m-3u ; d7=E=15m ; d6=5.4m=k ######### (d21 4u do 20u pl1 p1 ph10):f1 (d20 p14:sp3 ph0 d7 p14:sp5 ph0):f2 (d20 d8 p21*2 ph0 d7 d14 d8 d10):f3 ;######## end t2 ############## (p21 ph0):f3 4u p16:gp5 d16 (p11:sp1 ph3:r):f1 10u 20u pl1:f1 (p1 ph0):f1 3u p16:gp4 d16 4u d5 pl11:f1 ;d4 substituted by d5 DMO ;p11 ph4:r ;p2 substituted by p11 DMO (p11:sp1 ph4:r):f1 3u 20u pl1:f1 (d3 p1*2 ph5) (p21*2 ph0):f3 3u 20u pl11:f1 ; p11 ph4:r (p11:sp1 ph4:r):f1 3u d5 ;d4 substituted by d5 DMO p16:gp4 d16 pl16:f3 4u BLKGRAD go=2 ph31 cpd3:f3 d11 do:f3 mc #0 to 2 F1PH(ip1, id0 & dd9) F2PH(rd0 & rd9 & ip8, id10 & dd20) exit ph0=0 ph1=0 0 2 2 ph2=0 2 ph3=2 ph4=0 ph5=2 ph7=0 ph8=0 0 0 0 2 2 2 2 ph9=1 ph10=3 ph31=0 0 2 2 2 2 0 0 ;p34: 200(Gly)-325(Ala)-650us(Ser-Thr), f2 180 degree 13C shape ;sp20: f2 power 180 (CA) ;spnam20: Q3 gauss works fine ;spoffs20: 56ppm for HNN and HNN-Ala and 64ppm for HNN-ST ;d1: relaxation delay; 1-5*T1 ;d11: delay for disk I/O [30 msec] ;pl1: f1 power H high power pulse ;p1: f1 90 degree high power pl1 pulse ;d2: 2.25-2.75m (Jhn=92Hz) ;d3: p21-p1 (center pulse) ;pl2: f2 power C13 hard pulse (not used) ;pl3: f3 power N15 hard pulse ;p21: f3 90 degree high power N15 high power pl3 pulse ;pl11: f1 power H2O soft rectangular pulse (1m) ;p11: f1 90 degree H2O low power pl11 rectangluar pulse ;p13: 90 degree 13C shape ;p14: 180 degree 13C shape ;gpz1: 30% ;gpnam1: sine.100 shape of gradient pulse ;sp5: f2 power 180 (CA) soft shaped ;p14: f2 soft shaped pulse 180 null at 18200 Hz ;spnam5: use shape G3.256 ;spoff5: use offset 18200.0 Hz (600 MHz) on CO, null CA ;spnam3: use shape G3.256 ;spoff3: use offset 0.0 Hz (600 MHz) on CA, null CO ;d6: 5.4m (k in sequence) ;pl19: f1 power for cpds1 decoupling ;cpd1: dipsi2 (along x) ;pcpd3: f1 cpds1 pulse length (60-70u) use 65u ;d0: t1/2 ;d7: Tn [12-16m] use 15m ;d9: d0+d14+d7+d8 (Tn-t1/2) SET ALWAYS d9<=d8+3u ;gpz2: 30 % power level ;gpnam2: sine.100 shape of gradient pulse ;sp2: f2 power 90 (CA) soft shaped ;spnam2: use shape G4.256 ;spoff2: use offset 0.0 on CA, null CO ;sp4: f2 power 90 (CA) soft shaped ;spnam4: use shape G4.256tr (time reverse shape) ;spoff4: use offset 0.0 on CA, null CO ;d15: [12-18m] Tcn, use 16m ;gpz3: 30% ;gpnam3: sine.100 shape of gradient pulse ;d20: t2/2 (Tn-t2/2) [d9-d8] ;d10: t2/2 ;sp1: f1 power H2O soft shaped pulse ;p11: (2m) half-gaussian soft pulse with power sp1 ;spnam1: hg.1000 half gaussian shape ;p16: gradient pulse 1 ms ;gpz4: 80 % ;gpnam4: sine.100 shape of gradient pulse ;d4: 2.7m-p2-3u-4u-p16-d16 ;pl16: f3 power level N15 cpd decoupling ;cpd3: cpdprg3 decoupling sequence [garp] ;pcpd3: f3 decoupling 90 degree pulse [>170 u] ;l1: td1/2 STATES/TPPI ;l2: td2/2 STATES/TPPI ;in0: 1/(2* SW(N15)) = DW(N15) ;in9: SET ALWAYS in9=in0 ;nd0: 2 ;in10: 1/(2* SW(N15)) = DW(N15) ;in20: SET ALWAYS in20=in10 ;nd10: 2 ;NS: 8*n ;DS: 16 ;td1: number of experiments N15 ;td2: number of experiments N15 ;MC2: STATES-TPPI