#include "bits.nt" ;3D CBCACONH-J(CH) ;James Chou and Ad Bax Feb. 14,2001 ;JACS 2001, submitted ;Modified from the original pulse scheme by ;Grzesiek and Bax JACS 1992, 114, 6291-6293 ;This sequence does three 3D interleaved experiment to measure both CaHa and ;(CbHb2 + CbHb3) J coupling by quantitative J approach. The 3 instensities ;correspond respectively to 1.93, 3.73, and 7.22 ms of effective H-C J-modulation. ;The fitting program SFIT for calculating the couplings can also be downloaded from ;the same website. #define CARBON #define NITROGEN #define INTERLEAVE ;undefine this for just the reference spectrum define loopcounter NLOOP define loopcounter CLOOP "NLOOP=45" "CLOOP=61" ;All parameters for 600MHz spectrometer ;H = hydrogen frequency on water ;p1 = high power proton 90 at pl1 ;p30= low power proton 90 at pl30 (~15db) CPD decoupling cpds1 ;C1 = carbon frequency at 46 ppm, C2 = 177 ppm ;p4 =low power carbon 90 49us at pl4 ;p3 =22us 90 at pl3 for killing Ca/Cb boltzman magnetization ;p5 =22us at pl5, 2*22us 180 for C' refocusing ;p14 = 150 us sinc1.0 180 on C', sp1 (~10db) ;p16 = 500 us hsec.3 on 13C, sp2 (~3db) for C1 and sp0 for C2 ; for hsec.3, Q=|w(eff)/(d(theta)/dt)|=3 BW: 20kHz ;N = nitrogen frequency (~118 ppm) ;p7 = high power nitrogen 90 ~50us at pl7 ;p31 = low power 90 pulse ~200u at pl31 waltz16 cpd2 ;****** CA/CB evolution ****** ;in16 = in18 = 1/(2sw) ;MAX CLOOP = d18/in18 ;****** N evolution ****** ;in22=in24=in25=1/(4sw) ;MAX NLOOP = d22/in22 "d3=2.7m-p22" "d4=10u + p4*1.26" "d5=20u" ;universal dd, id, ip delay "d6=0.26m-p24" "d7=0.26m-p25" "d9=5.4m-p26" "d11=50m" ;universal dd, id, ip delay "d13=p16*0.5-p1" "d15=p4*1.26" "d20=1.8m-p1*4-6u" "d26=p7-p1" "d27=p16*0.5-p1*2-3u" "d28=400u-p14" "d29=p16*0.5-p7" ;**Increment or Decrement delays** "d16=10u" "d18=d16+3.4m" "d22=6.8m" "d24=d22-5.4m" "d25=d22" ;-------------- Gradient pulses ---------------------------- ;you have to set them by hand ;"p20=2.0m" ;gp0=(30%x,30%y) (sine.32) ;"p21=1.0m" ;gp21=50%x (sine.32) ;"p22=0.4m" ;gp3=50%x, gp3=50%y (sine.32) ;"p23=1.4m" ;gp23=(-30%y,-30%z) (sine.32) ;---The following must be optimized for different machine---- ;"p26=2.705m ;gp26=+50%z, gp27=-50%z (rectangular) ;"p24=0.2m ;gp24=+50%z (rectangular) ;"p25=0.074m ;gp25=-50%z (rectangular) ;****** end declaration ****** ze 1m ru1 2 5u do:N d11 d5 3 d5*4 4 d5*2 d5 5 d5*4 6 10u ;******* presaturation ******* 10u LOCK_ON 20u 10u pl3:C1 (p3 ph0):C1 ;kill Ca/Cb boltzman magnetization p21:gp21 50m pl1:H 1m pl6:C1 1m pl7:N d1 10u LOCK_OFF ;******* start 90-degree on ha/hb******* (p1 ph0) 1.3m (p16:sp2 ph0):C1 (d13 p1*2 ph0):H 1.3m (p1 ph7) 4u p20:gp0 300u pl4:C1 ;******* inept to c aliphatic ******* (p4 ph4):C1 6u (p16:sp2 ph0):C1 (d27 p1 ph0 3u p1*2 ph1 3u p1 ph0):H ;d27=p16*0.5-p1*2-3u 6u d15 ;p4*1.26 compensate for zero C evolution d16 ;d16=10u increment delay (p14:sp1 ph0):C2 ;sinc1.0 C' refocusing (~150u) if "l7==1" goto 71 ;1.93m length of effective J-modulation if "l7==2" goto 72 ;3.73m ... if "l7==3" goto 73 ;7.22m ... 71 d28 ;400u-p14 300u (p1 ph0 3u p1*2 ph1 3u p1 ph0):H d20 ;1.8m-p1*4-6u 900u goto 74 72 d28 ;400u-p14 1.2m (p1 ph0 3u p1*2 ph1 3u p1 ph0):H d20 ;1.8m-p1*4-6u goto 74 73 d28 ;400u-p14 d20 ;1.8m-p1*4-6u 1.2m (p1 ph0 3u p1*2 ph1 3u p1 ph0):H 74 4u (p16:sp2 ph17):C1 d18 pl4:C1 ;d18=d16+3.4m decrement delay 4u pl30:H ;******* inept to ca ******* (p4 ph19):C1 4u cpds1:H (p16:sp0 ph10):C2 ;hsec.3 compensation 5u (p16:sp2 ph29):C1 ;hsec.3 compensation d4 3.4m pl6:C1 (p16:sp0 ph10):C2 ;500u hsec.3 on C' 5u (p16:sp2 ph29):C1 ;500u hsec.3 on Ca 6u 3.4m pl4:C1 (p4 ph0):C1 10u 20u 10u pl15:C2 10u do:H p23:gp23 1m pl30:H 10u cpds1:H ;******* inept to carbonyl ******* (p15 ph8):C2 4.5m pl6:C1 (p16:sp2 ph29):C1 5.5m 5u 1m pl5:C2 (p5*2 ph10):C2 (p7*2 ph10):N 11m pl15:C2 (p16:sp2 ph29):C1 5u pl15:C2 (p15 ph18):C2 ;******* inept to nitrogen ******* (p7 ph27):N 11u pl4:C1 d22 pl3:C1 (p3*2 ph0):C1 8u pl6:C2 d22 (p16:sp0 ph0):C2 (d29 p7*2 ph27):N 8u pl3:C1 d25 (p3*2 ph0):C1 d24 3u do:H if "l1==1" goto 88 8u p26:gp26 ;Rance-Kay encoding gradient (+) d9 pl1:H (p7 ph19):N (d26 p1 ph0):H goto 89 88 8u do:H p26:gp27 ;Rance-Kay encoding gradient (-) d9 pl1:H (p7 ph9):N (d26 p1 ph0):H ;--------- end CT evolution on N -------------------- 89 2u p22:gp3 d3 (d26 p1*2 ph0):H (p7*2 ph27):N 2u p22:gp3 d3 (p1 ph15):H (p7 ph28):N 2u p22:gp4 d3 (d26 p1*2 ph0):H (p7*2 ph27):N 2u p22:gp4 d3 (d26 p1 ph0):H 4u p24:gp24 ;Rance-Kay decoding gradient (+) d6 (p1*2 ph0):H 2u p25:gp25 ;Rance-Kay decoding gradient (-) d7 pl31:N (2u ph0) go=2 ph31 cpd2:N 5u do:N d11 wr #0 if #0 zd 1m LOCK_ON ; #ifdef INTERLEAVE 10u iu7 lo to 3 times 3 10u ru7 #endif ; #ifdef NITROGEN 10u iu1 lo to 3 times 2 d12 dd22 10u ru1 d12 id24 d12 id25 lo to 4 times NLOOP ;46 ;change back to 46 d12*0.50 rd22 d12*0.25 rd24 d12*0.25 rd25 #endif ; #ifdef CARBON d5 ip4 lo to 5 times 2 d5 id16 d5 dd18 d5 ip31 d5 ip31 lo to 6 times CLOOP d5 rd16 d5 rd18 #endif ; 10u LOCK_ON 10u do:N exit ; ph0=0 ph1=1 ph2=2 ph4=0 ph7=1 3 ph27=0 ph8=0 0 0 0 2 2 2 2 ph28=1 ph9=2 ph10=0 ph12=0 ph15=1 ph16=3 ph17=0 0 1 1 2 2 3 3 ph18=(360) 0 ;adjusted for BS compensation ph19=0 ph20=0 ph29=0 ph31=0 2 2 0 2 0 0 2