;hcacongp3d-hdx.fm ;avance-version (12/01/11) ;HCA(CO)N experiment for the measurement of very fast backbone amide exchange ; based on publications by P. Peluppesy (Paris) ; ;options: ; no option CPMG experiment on 15N mixing in-phase and anti-phase ; LABEL_REF reference experiment with 1H decoupling switched on during CPMG ; to restrict relaxation completely to in-phase; set in ZG_OPTNS ; d25 is the time of the CPMG block where exchange takes place ; CNST50 is the number of pi pulses during d25; l4 is internally set to this ; ;3D sequence with ; inverse correlation for triple resonance using multiple ; inept transfer steps ; ; F1(H) -> F2(Ca) -> F2(C=O) -> F3(N, t1) ; -> F2(C=O) -> F2(Ca,t2) -> F1(H,t3) ; ;on/off resonance Ca and C=O pulses using shaped pulse ;phase sensitive (t1) ;phase sensitive using Echo/Antiecho gradient selection (t2) ;using constant time in t2 ; ;(V. Kanelis, L. Donaldson, D.R. Muhandiram, D. Rotin, ; J.D. Foreman-Kay & L.E. Kay, J. Biomol. NMR 16, 253-259 (2000)) ;(A.C. Wang, S. Grzesiek, R. Tschudin, P.J. Lodi & A. Bax, ; J. Biomol. NMR 5, 376-382 (1995)) ; ;$CLASS=HighRes ;$DIM=3D ;$TYPE= ;$SUBTYPE= ;$COMMENT= prosol relations= #include #include #include "p22=p21*2" "p2=p1*2" "d11=30m" "d4=1.6m" "d21=2.2m" "d22=4.5m" "d23=12m" "d24=d4*4/cnst11" "p30=5m" "d0=3u" "d10=d22/2" "d29=d22/2-d21" "d30=d22/2" "in0=inf1/2" "in10=inf2/4" "in29=in10" "in30=in10" "DELTA=d0*2+p8" "DELTA1=d22-d21-p14-4u" "DELTA2=d22-p14-4u" "DELTA3=d21-p16-d16-p26-4u" "DELTA4=p16+d16+4u" ;CPMG-block parameters "DELTA5=d25/(2*cnst50)-p22/2" "l4=cnst50" "spoff2=0" "spoff3=0" "spoff5=bf2*((cnst21-cnst22)/1000000)" "spoff8=0" "spoff9=0" "spoff13=bf2*((cnst26-cnst21)/1000000)" aqseq 321 1 d11 ze d11 pl12:f2 2 d11 do:f2 3 d1 pl1:f1 fq=cnst22(bf ppm):f2 (p1 ph1) d4 (center (p2 ph2) (p14:sp3 ph1):f2 ) d4 UNBLKGRAD (p1 ph2) (p13:sp2 ph1):f2 d21 pl19:f1 DELTA1 cpds1:f1 ph2 (p14:sp5 ph1):f2 4u (p24:sp9 ph4):f2 DELTA2 (p14:sp5 ph1):f2 4u (p13:sp8 ph1):f2 4u 30u fq=cnst21(bf ppm):f2 (p13:sp2 ph3):f2 d23 (center (p14:sp3 ph1):f2 (p22 ph1):f3 ) d23 (p13:sp8 ph2):f2 4u do:f1 (p26 ph10):f1 p16:gp3 d16 ; magnetization is two spin order 2C'zNz ; here comes the compensatory waltz16 block of duration d25 # ifdef LABEL_REF 20u d25 4u # else 20u cpds1:f1 ph1 d25 4u do:f1 # endif ; here ends the compensatory waltz16 block, and starts the CPMG block ; that either (1) mixes 2HzNxC'z antiphase in with in-phase NyC'z ; or (2) decouples 1H to maintain exclusively in-phase NyC'z (p21 ph1):f3 # ifdef LABEL_REF 20u cpds1:f1 ph1 4 DELTA5 (p22 ph2):f3 DELTA5 lo to 4 times l4 20u do:f1 # else 20u 5 DELTA5 (p22 ph2):f3 DELTA5 lo to 5 times l4 20u # endif /*LABEL_REF*/ (p21 ph1):f3 ; here ends the CPMG block and a clean up gradient is given p16:gp5 d16 (p26 ph2):f1 20u cpds1:f1 ph1 (p21 ph5):f3 d0 (p8:sp13 ph4):f2 d0 (p22 ph1):f3 DELTA (p21 ph1):f3 4u do:f1 (p26 ph9):f1 p30:gp4 d16 (p26 ph1):f1 20u cpds1:f1 ph2 (p13:sp2 ph1):f2 d23 (center (p14:sp3 ph1):f2 (p22 ph1):f3 ) d23 (p13:sp8 ph2):f2 4u 30u fq=cnst22(bf ppm):f2 (p13:sp2 ph6):f2 4u (p14:sp5 ph1):f2 d30 (p22 ph4):f3 d30 (p24:sp9 ph1):f2 4u (p14:sp5 ph1):f2 d10 (p22 ph4):f3 d29 4u do:f1 (p26 ph10):f1 p16:gp1*EA d16 DELTA3 pl1:f1 (center (p1 ph1) (p13:sp8 ph7):f2 ) d24 (center (p2 ph1) (p14:sp3 ph1):f2 ) d24 (center (p1 ph2) (p13:sp2 ph8):f2 ) d4 (center (p2 ph1) (p14:sp3 ph1):f2 ) d4 (p1 ph1) DELTA4 (p2 ph1) p16:gp2 d16 pl12:f2 4u BLKGRAD go=2 ph31 cpd2:f2 d11 do:f2 mc #0 to 2 F1PH(calph(ph5, +90), caldel(d0, +in0)) F2EA(calgrad(EA) & calph(ph8, +180), caldel(d10, +in10) & caldel(d29, +in29) & caldel(d30, -in30) & calph(ph6, +180) & calph(ph31, +180)) exit ph1=0 ph2=1 ph3=0 0 0 0 2 2 2 2 ph4=0 0 0 0 0 0 0 0 2 2 2 2 2 2 2 2 ph5=0 2 ph6=0 ph7=0 0 2 2 ph8=3 3 1 1 ph9=3 ph10=2 ph31=0 2 2 0 2 0 0 2 ;pl1 : f1 channel - power level for pulse (default) ;pl3 : f3 channel - power level for pulse (default) ;pl12: f2 channel - power level for CPD/BB decoupling ;pl19: f1 channel - power level for CPD/BB decoupling ;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) ;sp8 : f2 channel - shaped pulse 90 degree (on resonance) ; sp8 for time reversed Calpha pulses ;sp9 : f2 channel - shaped pulse 180 degree (Ca on resonance) ; sp9 requires higher selectivity than sp3 (Ca only) ;sp13: f2 channel - shaped pulse 180 degree (Ca and C=O, adiabatic) ;p1 : f1 channel - 90 degree high power pulse ;p2 : f1 channel - 180 degree high power pulse ;p8 : f2 channel - 180 degree shaped pulse (Ca and C=O, adiabatic) ;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 ;p30: gradient pulse 2 [5 msec] ;d0 : incremented delay (F1 in 3D) [3 usec] ;d1 : relaxation delay; 1-5 * T1 ;d4 : 1/(4J(HCa)) [1.6 msec] ;d10: incremented delay (F2 in 3D) = d22/2 ;d11: delay for disk I/O [30 msec] ;d16: delay for homospoil/gradient recovery ;d21: 1/(3J(HCa)) [2.2 msec] ;d22: 1/(4J(CaCO)) [4.5 msec] ;d23: 1/(4J(NCa)) [12 msec] ;d24: 1/(8J(HCa)) for all multiplicities ; 1/(4J(HCa)) for CH ;d25: length of total CPMG period [20 msec] ;d29: incremented delay (F2 in 3D) = d22/2-d21 ;d30: decremented delay (F2 in 3D) = d22/2 ;cnst11: for multiplicity selection = 8 for all multiplicities, 4 for CH ;cnst21: CO chemical shift (offset, in ppm) ;cnst22: Calpha chemical shift (offset, in ppm) ;cnst26: Call chemical shift (offset, in ppm) [101 ppm] ;cnst50: Number of 15N 180 pulses during spin lock [2,4,8,16] ;o2p: Calpha chemical shift (cnst22) ;inf1: 1/SW(N) = 2 * DW(N) ;inf2: 1/SW(Ca) = 2 * DW(Ca) ;in0: 1/(2 * SW(N)) = DW(N) ;nd0: 2 ;in10: 1/(4 * SW(Ca)) = (1/2) DW(Ca) ;nd10: 4 ;in29: = in10 ;in30: = in10 ;ns: 8 * n ;ds: 32 ;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: echo-antiecho in F2 ;cpds1: decoupling according to sequence defined by cpdprg1 ;cpd2: decoupling according to sequence defined by cpdprg2 ;pcpd1: f1 channel - 90 degree pulse for decoupling sequence ;pcpd2: f2 channel - 90 degree pulse for decoupling sequence ;zgoptns: specify ZGOPTNS = -DLABEL_REF for 1H decoupling during CPMG ;use gradient ratio: gp 1 : gp 2 : gp 3 : gp 4 : gp5 ; 80 : 20.1 : 40 : 60 : 5 ;for z-only gradients: ;gpz1: 80% ;gpz2: 20.1% ;gpz3: 40% ;gpz4: 60% ;gpz5: 5% ;use gradient files: ;gpnam1: SMSQ10.100 ;gpnam2: SMSQ10.100 ;gpnam3: SMSQ10.100 ;gpnam4: SMSQ10.100 ;gpnam5: SMSQ10.100 ;preprocessor-flags-start ;LABEL_REF: when this is defined in ZGOPTNS then a referennce experiment is executed ; option -DLABEL_REF (eda: ZGOPTNS) ;$Id: hcacongp3d,v 1.5.8.1 2012/01/31 17:56:22 ber Exp $