;DFtstrhsqcnoesyhsqcsi.2 ;avance-version (05/26/05) ;TIME-SHARED HN/HC-HSQC-NOESY-HC-PEP-HSQC/HN-TROSY ;gives four 4Ds in one experiment: need processing scripts: extract4D and add.com ;developed for ILV-1H-Me samples (otherwise deuterated): ; use zgoption LABEL_CN ; ; ; ;4D sequence with ; homonuclear correlation via nOe ; correlation may be due to noe or chemical exchange ; Simultaneous H-1/X correlation via time shared inepts or PEP/ST2-PT transfers: ; |HN ->N(t2)->HN(t1)->HN->N(t3)->HN(t4) ; |HN ->N(t2)->HN(t1)->HMe->C(t3)->HMe(t4) ; |HMe ->C(t2)->HMe(t1)->HN->N(t3)->HN(t4) ; |HMe ->C(t2)->HMe(t1)->HMe->C(t3)->HMe(t4) ; ; ********************************************* ; * allows to fold HMe region on HN region * ; *high resolution Hindirect in t1 * ; * separation and unfolding during processing* ; ********************************************* ; ;using sensitivity improvement for 13CMe/HMe and TROSY for 15N/HN ;phase sensitive (t1) ;phase sensitive (t2) ;phase sensitive using Echo/Antiecho-TPPI gradient selection (t3) both N and C ;with decoupling during acquisition on 13C ;using flip-back pulse ;************************************************************************ ;(use parameterset NOESYHSQCFPf3GPSI3D), then addapt as follows: ; ;cnst10: offset (ppm) for shaped inversion pulse on HMe ; have tested : ; cnst10=1: REBURP on HMe (1ppm): OK but 15N relaxation ; cnst10=-2: Q3 on Hali (-2ppm): 600us at 750 MHz (7.56 ppm bwidth) ;cnst11: offset for HMe 90 pulse (1ppm) ; warning: depending on wether water is touched or not ; you will need to add flip-backs as indicated in comments ; in the pulse program ;cnst13: 15N spectral width in ppm ;cnst17: center of nOe region: fold HMe on HN ; cnst17=8 to fold Me on HN, SW(Hindirect)= c.a. 5ppm ; cnst17=1 to fold HN on HMe, SW(Hindirect)= c.a. 5ppm ;cnst18: =o1p: water ;PROSOL compatible, but : then update the two following pulses: ;p27: TRIM pulse ;p28: HMe 90 pulse ;p29: HMe inversion pulse ;sp8: HMe 90 ;sp9: HMe inversion ;spnam8: 90 HMe: Sinc1.1000 or Q5 or... ;spnam9: 180 HMe, Q3 (or REB) ;Frueh DP, Vosburg DA, Walsh CT, Wagner G, JBNMR 2006 (In press 01/05/06) ;4D TROSY TS Dfrueh 052605 prosol relations= #include #include #include "p2=p1*2" "p22=p21*2" "d0=3u" "d2=1s/(cnst3*4)-p16-d16" "d3=1s/(cnst2*2)" "d4=1s/(cnst3*2)-1/(cnst2*2)-p3-p16-d16" "d10=3u" "d20=3u" "d6=1s/(cnst2*4)-p4/2-p16-d16" "d5=1s/(cnst3*4)-1s/(cnst2*4)-p4/2" "d21=3u" "d22=3u" "d11=30m" "d12=20u" "d14=1s/(cnst3*4)-p16-d16-p11-15u" "d15=1s/(cnst3*4)-1s/(cnst2*4)-p4/2-p11-15u" "d13=4u" ;"d17=d16-p28/4" "d17=d16" "d18=d8-d3-d4-3*p16-2*d16-2*p3 -p21-p11" "d26=1s/(cnst3*4)" ;1/4JNH "d24=1s/(cnst2*4)" ;1/4JCH "d23=d24-p4/2" ;1/4JCH "d25=d26-d24-p4/2" ; "p19=300u" "DELTA1=d13+p19+d16+4u-p28/2" "DELTA3=d26-p16-d16" "DELTA4=d26-p16-d16" "DELTA5=d10" "DELTA7=d20+p29+p3+2*p16+2*d16+2*d10" "DELTA8=d20+p29+2*d10" "DELTA9=(d26+d24)/2" "DELTA10=(d26-d24)/2" "DELTA11=(d26+d24)/2+p22/2" "DELTA12=(d26-d24)/2-p22/2" "DELTA13=(d26-d24)/2-p28/2" ;"DELTA13=(d26-d24)/2" "DELTA16=d3-p11" "DELTA14=2*d10+p2" "DELTA15=2*(2*d10+p2)+2*p3+2*d20+p4" ;# ifdef LABEL_CN "DELTA=p16+d16+larger(p2,p4)+d10*2" "DELTA2=larger(p4,p22)+d0*2" ;# else ;"DELTA=p16+d16+p2+d10*2" ;"DELTA2=p22+d0*2" ;# endif /*LABEL_CN*/ "TAU=d8-p16-d16-p19-d16-10u" "in20=1/(2*cnst13*bf3/1000000)-in10" "in21=in10" "in22=in20" "spoff9=bf1*((cnst10)/1000000)-o1" "spoff8=bf1*((cnst11)/1000000)-o1" "l0=td1/2" ;"l10=sqrt(td2)" aqseq 321 1 ze d11 pl12:f2 2 d11 do:f2 3 d12 pl1:f1 fq=cnst18(bf ppm):f1 ;H2O d1 pl3:f3 pl2:f2 5u 5u pl0:f1 5u (p11:sp1 ph23):f1 5u 5u pl1:f1 5u ;start 1st HSQC (p1 ph1):f1 d2 p16:gp7 d16 (center (p22 ph1):f3 (p2 ph1):f1) d5 (p4 ph1):f2 d6 p16:gp7 d16 (p27 ph1):f1 4u (p1 ph21):f1 5u ; 5u pl0:f1 ; 5u ; (p11:sp1 ph21:r):f1 ; 5u p16:gp8 d16 pl1:f1 (p21 ph13):f3 ;N========== DELTA15 (p22 ph2):f3 d22 ; here we save relaxation for 13C (1H Z) (p3 ph3):f2 ;CN DELTA14 ;for zero phase (p4 ph1):f2 ;for zero phase d21 (p2 ph22):f1 d21 (p3 ph1):f2 d22 (p21 ph1):f3 p16:gp9 d16 5u 5u pl0:f1 5u (p11:sp1 ph23):f1 5u 5u pl1:f1 5u (p1 ph1):f1 4u d6 p16:gp11 d16 pl0:f1 (p4 ph1):f2 d15 (p11:sp1 ph22:r):f1 5u 5u pl1:f1 5u (center (p22 ph1):f3 (p2 ph1):f1) 5u 5u pl0:f1 5u (p11:sp1 ph22:r):f1 d14 p16:gp11 d16 pl1:f1 (p1 ph21):f1 ; NOESY--------- 5u 5u pl0:f1 5u (p11:sp1 ph23:r):f1 ;water restored to Z p16:gp17 d16 pl1:f1 5u (p11:sp1 ph6:r):f1 ;to compensate next 90 5u d16 pl1:f1 5u d12 fq=cnst17(bf ppm):f1 ;center NOE 5u (p1 ph8) DELTA2 pl2:f2 (p2 ph9) d0 (center (p4 ph1):f2 (p22 ph1):f3 ) d0 (p1 ph10) ;Tm 5u pl0:f1 5u fq=cnst18(bf ppm):f1 ;H2O 5u (p11:sp1 ph18):f1 DELTA16 ;d3 (p3 ph1):f2 p16:gp12 d16 d4 (p21 ph1):f3 p16:gp14 d16 d18 5u 5u pl0:f1 5u (p11:sp1 ph12):f1 5u 5u pl1:f1 5u (p1 ph11);-----------2nd-HSQC----- d23 (p4 ph1):f2 d25 (center (p2 ph1) (p22 ph1):f3 ) d26 ;pl10:f1 ;(p27 ph11) ;5u pl1:f1 (p1 ph2) 4u pl0:f1 ; (p11:sp1 ph12):f1 4u pl3:f3 p16:gp2 d16 pl1:f1 (p21 ph4):f3 ;TShared N DELTA7 DELTA8 (p22 ph1):f3 d20 d20 (p3 ph14):f2 ;C p16:gp3*EA d16 DELTA5 (p29:sp9 ph22):f1 ;REB on Hc-Me: 4ppm @ 1 ppm DELTA5 (p4 ph1):f2 d10 (p29:sp9 ph7):f1 d10 p16:gp13*EA d16 (center (p1 ph17) (p3 ph16):f2 ) DELTA11 pl3:f3 (p4 ph1):f2 DELTA12 (center (p2 ph1) (p22 ph1):f3 ) DELTA9 (p4 ph1):f2 DELTA10 (center (p1 ph1) (p21 ph2):f3 (p3 ph2):f2 ) DELTA11 pl3:f3 (p4 ph1):f2 DELTA12 (center (p2 ph1) (p22 ph1):f3 ) DELTA9 (p4 ph1):f2 DELTA13 (center (p28:sp8 ph2):f1 (p21 ph16):f3) ;(p21 ph1):f3 DELTA1 (p2 ph1) d13 p19:gp6 d17 pl12:f2 4u BLKGRAD go=2 ph31 cpd2:f2 d11 do:f2 wr #0 if #0 zd 50u ip13*2 50u ip14*2 lo to 3 times 2 ;interleave for pathways editing HN->HN, HN->HMe, etc 50u igrad EA 50u ip16*2 50u ip17*2 50u ip12*2 lo to 3 times 2 50u id10 50u id20 50u ip4*2 50u ip14*2 50u ip31*2 lo to 3 times l20 ;l20 EA 50u rp12 50u rd10 50u rd20 50u ip3 50u ip13 50u rp4 50u rp14 lo to 3 times 2 50u ip31*2 50u id21 50u id22 lo to 3 times l10 ;l10: States-TPPI dim (t1 50u rp13 50u rp3 50u rd21 50u rd22 50u rp31 50u ip8 50u ip6 50u ip9 lo to 3 times 2 50u id0 lo to 3 times l0 ;l0 : 1H d11 do:f2 ;1D/2D/3D versions for testing ;mc #0 to 2 ; F1PH(rd10 & rd20 & rd21 & rd22 &ip8 & ip9, id0) ;test H ; F2EA(igrad EA & ip16*2 & ip17*2, id10 &id20 & ip6*2) ;test EA ; F2PH(ip3 & ip13, id21 &id22) ;test St-TPPI exit ph1=0 ph2=1 ph3=0 2 ph4=0 0 2 2 ph5=1 1 ;3 3 ph15=3 3 ;1 1 ph6=2 2 2 2 0 0 0 0 ph7=0 0 2 2 ph8=0 0 0 0 2 2 2 2 ph9=1 1 1 1 3 3 3 3 ph10=0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 ph18=2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ph11=0 0 0 0 0 0 0 0 2 2 2 2 2 2 2 2 ph12=2 2 2 2 2 2 2 2 0 0 0 0 0 0 0 0 ph13=0 2 ph14=0 0 2 2 ph16=0 ph17=1 ph20=0 ph21=1 ph22=2 ph23=3 ph25=3 ph31=0 2 2 0 2 0 0 2 2 0 0 2 0 2 2 0 2 0 0 2 0 2 2 0 0 2 2 0 2 0 0 2 ;pl0 : 120dB ;pl1 : f1 channel - power level for pulse (default) ;pl3 : f3 channel - power level for pulse (default) ;sp1 : f1 channel - shaped pulse 90 degree ;p1 : f1 channel - 90 degree high power pulse ;p2 : f1 channel - 180 degree high power pulse ;p11: f1 channel - 90 degree shaped pulse ;p16: homospoil/gradient pulse [1 msec] ;p19: homospoil/gradient pulse [300 usec] ;p21: f3 channel - 90 degree high power pulse ;p22: f3 channel - 180 degree high power pulse ;d0 : incremented delay (F1 in 3D) [3 usec] ;d1 : relaxation delay; 1-5 * T1 ;d8 : mixing time ;d10: incremented delay (f3 in 3D) [3 usec] ;d11: delay for disk I/O [30 msec] ;d12: delay for power switching [20 usec] ;d13: short delay [4 usec] ;d16: delay for homospoil/gradient recovery ;d24: 1/(4J)CH for CH ; 1/(8J)CH for all multiplicities ;d26: 1/(4J(NH)) ;cnst2: = J(CH) ;cnst3: = J(NH) ;in0: 1/(2 * SW(H)) = DW(H) ;nd0: 2 ;in10: 1/(2 * SW(C)) = DW(C) ;nd10: 2 ;NS: 8 * n ;DS: >= 16 ;td1: number of experiments in F1 ;td2: number of experiments in f3 ;FnMODE: States-TPPI (or TPPI) in F1 ;FnMODE: echo-antiecho in f3 ;cpd2: decoupling according to sequence defined by cpdprg2 ;pcpd2: f2 channel - 90 degree pulse for decoupling sequence ;l10: complex States-TPPI N/C: l10*l20=td2/8 ;l20: complex E/A N/C: l10*l20=td2/8 ;use gradient ratio: gpz3=-3(p19)(gpz6)/p16 ; gpz13=-7(p19)(gpz6)/p16 ; ;for z-only gradients: ;gpz1: 30% ;gpz2: -50% ;gpz3: -23.9% ;gpz6: 27% ;gpz7: 40% ;gpz8: 37% ;gpz9: -63% ;gpz11: 13% ;gpz12: 7% ;gpz13: -56.1% ;gpz14: 11% ;gpz17: -63 ;use gradient files: ;gpnam1: SINE.100 ;gpnam2: SINE.100 ;gpnam3: SINE.100 ;gpnam6: SINE.50 ;gpnam7: SINE.100 ;gpnam8: SINE.100 ;gpnam9: SINE.100 ;gpnam17: SINE.100 ;gpnam12: SINE.100 ;gpnam13: SINE.100 ;gpnam14: SINE.100 ;preprocessor-flags-start ;LABEL_CN: for C-13 and N-15 labeled samples start experiment with ; option -DLABEL_CN (eda: ZGOPTNS) ;preprocessor-flags-end ;modified from: $Id: noesyhsqcfpf3gpsi3d,v 1.1.2.3 2003/06/18 15:27:56 ber Exp $