#include "bits.nt" ;3D TROSY-HNCOJ ;3D interleaved TROSY-HNCO for measurement of J(NC') + D(NC') ;James Chou and Ad Bax July 12, 2000 ; ;Citaion: ; James Chou, Frank Delaglio and Ad Bax ; J. Biomol NMR, 18, 101-105, 2000. ;--RF Pulses--------------- ;p1 = 90 deg (10us) 1H pulse @pl1 ;p2 = 90 deg @pl2 1H; should be ~1ms ;p3 = 90 hard on C' ;p6 = sp0 90 sinc1.0 on C' (~100 us at 800 MHz), center lobe sinx/x ;p7 = 90 deg (~50us) 15N pulse @pl7 ;p8 = sp3 sinc1.0 on CA (~100 us at 800 MHz) ;p16= sp2 180 hsec.3 on C' (~600-1000 us at 800 MHz), hyperbolic secant ;--Gradients--------------- ;p21=1m, gp1=5%x sine.50 ;sine bell shaped gradient ;p20=1m, gp0=30%y sine.50 ;p20=1m, gp2=30%x sine.50 ;p21=1m, gp3=30%z sine.50 ;--Options----------------- #define NITRO #define CARBON #define INTERLEAVE ;--Evolutions-------------- define loopcounter NLOOP define loopcounter CLOOP "NLOOP=60" ;max = 62 for 500u increment in 15N dim "CLOOP=32" ;number of complex C' increments ; ;15N evolution in20=in21=in22 Total evolution = 4*in20*NLOOP ;C' evolution in0 Total evolution = 2*in0*CLOOP ;--Delays------------------ "d0=in0*0.5-p7-p6*0.54" ;90,-180 phase correction in C' dim "d3=2.5m-p21" "d4=2.6m-p21-p2" "d11=50m" "d16=p16" "d19=in21*0.444" ;fudge delay for zero ph1 in 15N dim "d20=7.5m+p16*0.5" "d21=7.5m" "d22=7.5m-p2-16u" "d26=p7-p1" ; 1 ze 1m RESET 2 d11 do:N 3 3m 7 5u do:C1 10u pl3:C1 d1 LOCK_ON 1m LOCK_OFF (p3 ph0):C1 ;kill C' boltzman magnetization 10u pl1:H 10u pl7:N ;------- start 90-degree on 1HN ----------------------- (p1 ph0):H ;the first 1H 90 3u p21:gp1 ;1m @ 5%x d3 (d26 p1*2 ph0):H (p7*2 ph0):N p21:gp1 ;1m @ 5%x d3 (p1 ph1):H 3u ;------- INEPT to 15N ----------------------- (p7 ph0):N ;INEPT to 15N if "l2==2" goto 20 ;interleaving expt with 33 and 67msec J(NC') dephasing 16.6m pl6:C1 (p16:sp2 ph0):C1 ;selective C' adiabatic 180 16.6m goto 21 20 33.2m pl6:C1 (p16:sp2 ph0):C1 21 (p7*2 ph0):N d16 ;d16 = p16 33.2m (p7 ph1):N p20:gp0 ;scrambler gradient 1m @ 30%y 1m pl5:C2 (p6:sp0 ph6):C1 ;selective C' 90 (sinc1.0) ;------ Start C' evolution----------- d0 ;start C' evolution (p8:sp3 ph0):C1 (p7*2 ph0):N ;CA and 15N refocusing d0 ;------ End C' evolution------------- (p6:sp0 ph0):C1 ;selective C' 90 (sinc1.0) 6u p20:gp2 ;scrambler gradient 1m @ 30%x 1m if "l1==1" goto 88 ;select appropriate TROSY phase (p7 ph17):N ;ph17=1 2 --| goto 89 ; | 88 (p7 ph7):N ;ph7=1 0 --| ;------ Start CT 15N evolution------- 89 d20 ;decrement d20 = 7.5m (p8:sp3 ph0):C1 d20 ;decrement d20 = 7.5m (p7*2 ph0):N (p16:sp2 ph0):C1 d19 ;fudge delay for zero ph1 in 15N dim d21 ;increment d21 (p8:sp3 ph0):C1 d22 ;increment d22 5u pl2:H (p2 ph0):H ;1m water 90 pulse; ST2 transfer 6u 5u pl1:H ;------ End CT 15N evolution--------- if "l1==1" goto 78 ;select appropriate TROSY phase (p1 ph12):H ;ph12 = 1 --| goto 79 ; | 78 (p1 ph2):H ;ph2 = 3 --| 79 5u p21:gp1 ;1m @ 5%x d3 (d26 p1*2 ph0):H (p7*2 ph0):N 5u p21:gp1 ;1m @ 5%x d3 if "l1==1" goto 98 ;select appropriate TROSY phase (p1 ph0):H (p7 ph14):N ;DOUBLE 90 ph14 = 1 --| goto 99 ; | 98 (p1 ph0):H (p7 ph4):N ;DOUBLE 90 ph4 = 3 --| 99 5u p21:gp3 ;1m @ 30%z d4 pl2:H (p2 ph15):H 3u 5u pl1:H (d26 p1*2 ph0):H (p7*2 ph0):N 8u pl2:H (p2 ph15):H 5u p21:gp3 ;1m @ 30%z d4 pl30:C1 (p7 ph0):N go=2 ph31 cpd1:C1 ; CPD for C' during acquisition 10u do:C1 1m LOCK_ON d11 wr #0 if #0 zd ; #ifdef INTERLEAVE 10u iu2 ;increment loopcounter l2 lo to 3 times 2 ;reset loopcounter l2 to 1 10u ru2 #endif ; #ifdef NITRO 0.1m iu1 ;increment loopcounter l1 lo to 3 times 2 10u dd20 10u id21 10u id22 10u ru2 10u ru1 ;reset loopcounter l1 to 1 lo to 3 times NLOOP 10u rd21 10u rd22 10u rd20 #endif ; #ifdef CARBON 10u ip6 lo to 3 times 2 10u id0 10u ip31 ;TPPI-States in C' dim 10u ip31 lo to 3 times CLOOP #endif ; 1m do:C1 1m do:N 1m RESET 1m exit ph0=0 ph1=1 ph2=3 ph12=1 ph4=3 ph14=1 ph15=(360)178 ph6=0 0 2 2 ph7=1 0 ph17=1 2 ph10=(360)0 ph11=0 ph31=1 2 3 0 ;receiver phase