;cbcaconhse.ref ;3D CBCA(CO)NH - Gradient enhanced. ;Grzesiek and Bax, J. Am. Chem. Soc., 114, 6291-6293 (1992) ;Muhandiran and Kay, J. Magn. Reson. B 103, 203-216 (1994) ;Bruker Avance/Xwin-nmr version ;Written up by F. Abildgaard, NMRFAM (abild@nmrfam.wisc.edu) ;uthscsa avance600, aph, 06/02/02 ; ;NOTE1 - Follow the guidelines below for setting up this experiment ;Unless you fully understand what you are doing, do NOT proceed beyond ;the point indicated ;NOTE2 - In the header section that follows, anything between double quotes ;is something you can type at the XWINNMR command line ;NOTE3 - Anything followed by a "s" means a time in seconds, anything followed ;by a "m" means a time in milliseconds, and anything followed by a "u" ;means a time in microseconds ; ;Set the options below to tailor the pulse program for your needs ;#define ONE_D ;uncomment for 1D expt ;#define N15_EVOL ;uncomment for 3D and H1-N15 2D exp #define C13_EVOL ;uncomment for 3D and H1-C13 2D exp ;#define SHORT_P2 ;uncomment if p8/2 > p2 ; ;Set the channel assignments to match your hardware settings in "edasp" #define H f1 #define C f2 #define N f3 ;1H pulses - set carrier to H2O (ca 4.7 ppm) ;p1 - square 90 1H @ pl1, for excitation, typically p1 = 8u @ pl1 = -1dB ;pcpd1 - square 90 1H @ pl11, for cpd, typically pcpd1 = 45u @ pl11 = +15dB ; "cpdprg1 dipsi2" ;15N pulses - set carrier to amide center (ca 118 ppm) ;p2 - square 90 15N @ pl2, for excitation, typically p2 = 45u @ pl2 = -4.4dB ;pcpd2 - square 90 15N @ pl12, for cpd, typically pcpd2 = 180u @ pl12 = +7.9dB ; "cpdprg2 waltz16" ;13C pulses - carrier is set according to fqlist3 frequency list ; chose/edit in ased, should have two values, ; first is CA/CB (43 ppm), second is CA (56 ppm) ;p3 - square 90 @ pl3, for Ca/Cb excitation, p3 must equal 48.2u,typically pl3= +7.8dB ;p6 - square 90 @ pl6, for Ca/Cb inversion, p6 must equal 21.6u,typically pl6= +0.9dB ;p4 - square 90 @ pl4, for Ca excitation, p4 must equal 53.5u, typically pl4 = +8.7dB ;p5 - shaped 180 @ sp5, for CO inversion, typically p5 = 250u @ sp5 = +9.5dB ; "spnam5 seduce1.jc", "spoffs5 20068" ;p7 - shaped 90 @ sp7, for CO excitation, p7 must equal 53.5u, typically sp7 = +8.6dB ; "spnam7 rect32", "spoffs7 18107" ;p8 - shaped 180 @ sp8, for CO inversion, p8 must equal 47.8u, typically sp8 = +1.5dB ; "spnam8 rect32", "spoffs8 18107" ;pcpd3 - shaped 90 @ sp3, for CA decoupling, typically pcpd3 = 125u @ sp3 = +9.5dB ; "spnam3 seduce1.jc", "spoffs3 0", "cpdprg3 waltz16sp3" ;Other settings ;d1 - recylcle time, typically d1 = 1s ;ns - n*4 where n is a positive integer ;ds - n*4 where n is a positive integer ;Gradient settings ;Note1 - grad times are preset, you do not need to adjust these ;Note2 - set grad params according to table below, note integer in first ;column is substituted to yield the parameter that is to be adjusted ;example for grad# 0, set "gpnam0 sine.64", "gpx0 0", "gpy0 0", "gpz0 12" ; ;grad# gpnam# gpx# gpy# gpz# ;0 sine.64 0 0 12 ;1 sine.100 0 0 29 ;2 sine.32 0 0 29 ;3 sine.100 0 0 20 ;4 sine.100 77 0 44 ;5 sine.20 77 0 44 ; ;N15 evolution ;in10 - incremented delay, set to 1/(2*SW), typically 300u ;in30 - incremented delay, set to 1/(2*SW), typically 300u ;l4 - number of complex points, maximum is trunc(d10/in10), typically l4 = 41 ;N15 Conversion/Processing ;Conversion, Dim=y, yMODE Echo-AntiEcho, aq2D States ;Example processing... ;| nmrPipe -fn SP -off 0.48 -end 0.98 -pow 2 -c 0.5 \ ;| nmrPipe -fn ZF -size 512 \ ;| nmrPipe -fn FT -neg \ ;| nmrPipe -fn PS -p0 0.0 -p1 0.0 -di -verb \ ;| nmrPipe -fn POLY -auto -ord 1 \ ;C13 evolution ;in0 - incremented delay, set to 1/(2*SW), typically 49.5u ;in20 - incremented delay, set to 1/(2*SW), typically 49.5u ;l6 - number of complex points, maximum is trunc(d20/in20), typically l6 = 57 ;C13 Conversion/Processing ;Conversion, Dim=z, yMODE Complex, aq2D States ;Example processing... ;| nmrPipe -fn SP -off 0.48 -end 0.98 -pow 2 -c 0.5 \ ;| nmrPipe -fn ZF -size 512 \ ;| nmrPipe -fn FT \ ;| nmrPipe -fn PS -p0 0.0 -p1 0.0 -di -verb \ ;| nmrPipe -fn POLY -auto -ord 1 \ ; ;Testing ;The 1H/15N plane can be tested by commenting N15_EVOL in and C13_EVOL out, strong signal ;The 1H/13C plane can be tested by commenting C13_EVOL in and N15_EVOL out, strong signal ;PRESET DELAYS - DO NOT ADJUST ANYTHING BEYOND THIS POINT ; ;gradients "p15=500u" "p16=1000u" "p17=250u" "p18=1.25m" "p19=125u" "p20=1.5m" ;sanity checks ; #ifdef ONE_D #undef N15_EVOL #undef C13_EVOL #endif ; aqseq 321 ; define delay TAUA define delay TAUB define delay TAUC define delay TAUD define delay TAUE define delay TAUF define delay TAUG define delay TAUH define delay T define delay C_H_N define delay C_H_CAB define delay C_N_CO define pulse H1_180 define pulse CAB_180 define pulse CA_180 define pulse CO_180 define pulse N15_180 "d0=3u" "d10=12.4m" "d30=12.4m-5.5m-d13*2" "d6=p1" "d7=p2" "d11=100m" "d12=10u" "d13=5u" "d14=40u" "d16=200u" "d17=50u" "d23=d13*2+d12" "d24=p4*2" "d20=3.6m-p5-p17-d16-d13-d12" "TAUA=1.8m-p15-d13" "TAUB=1.1m-p5-p17-d16-d13" "TAUC=3.6m-p5-d13*3-d12*2" "TAUD=4.4m" "TAUE=12.4m" "TAUF=5.5m-p18-d12-d13*2-d16" "TAUG=2.3m-p15" "TAUH=500u" "d18=TAUH-p19-d13" "T=d20" "H1_180=p1*2" "CAB_180=p6*2" "CA_180=p4*2" "CO_180=p8" "N15_180=p2*2" "C_H_CAB=p6-p1" "C_H_N=p2-p1" #ifdef SHORT_P2 "C_N_CO=p8*0.5-p2" #endif #define SED_ON d13 \n d12 pl0:C \n d13 cpds3:C #define SED_OFF d13 do:C #define H1_DEC_ON d13 \n d12 pl11:H \n d13 cpds1:H #define H1_DEC_OFF d13 do:H \n d12 pl1:H #include #include 1 ze 2 d11 do:N d14 d14 3 d14 d14 d14 d14 d14 d14 4 d14 d14 d14 5 d14 d14 d14 d14 6 20u LOCKH_OFF d1 pl1:H pl2:N ;--------------------- carbon freq. on Cab --------------- 50u fq3:C (p3 ph0):C 20u LOCKH_ON d13 p15:gp0 ;500u, 8G/cm d16 ;--------------------- INEPT to Cab ---------------------- (p1 ph0):H d13 p15:gp0 ;500u, 8G/cm TAUA pl6:C (C_H_CAB H1_180 ph0):H (CAB_180 ph0):C d13 p15:gp0 ;500u, 8G/cm TAUA (p1*55 ph0):H d13 (p1 ph1):H d13 p16:gp1 ;1m, 20G/cm d16 pl3:C (p3 ph11):C ;--------------------- Constant time Cab evolution -------- d0 d12 pl0:C (p5:sp5 ph0):C (N15_180 ph0):N d13 p17:gp2 ;250u, 20G/cm d16 (T) (TAUB H1_180 ph0):H d12 pl6:C (CAB_180 ph0):C 3u d20 p17:gp2 ;250u, 20G/cm d16 d12 pl0:C (p5:sp5 ph0):C d13 d12 pl3:C ;----------------------- INEPT to C' --------------------- (p3 ph0):C H1_DEC_ON d12 pl0:C (p5:sp5 ph0):C d13 TAUC pl6:C (CAB_180 ph12):C d12 pl0:C (p5:sp5 ph0):C d13 TAUC pl3:C d23 (p3 ph0):C H1_DEC_OFF p16:gp1 ;1m, 15G/cm d17 H1_DEC_ON ;----------------------- Switch frequency to Ca -------------- d16 fq3:C ;----------------------- INEPT to N -------------------------- d12 pl0:C (p7:sp7 ph13):C d13 d12 pl4:C (TAUE) (TAUD CA_180 ph0):C d12 pl0:C d24 (CO_180:sp8 ph0):C (TAUE) (N15_180 ph0):N d12 pl4:C (CA_180 ph0):C d13 d12 pl0:C (p7:sp7 ph0):C H1_DEC_OFF p20:gp3 ;1.5m, 20G/cm d17 H1_DEC_ON d16 pl0:C SED_ON (p2 ph14):N d10 SED_OFF d12 pl0:C #ifdef SHORT_P2 (CO_180:sp8 ph0):C (C_N_CO N15_180 ph15):N #else (CO_180:sp8 ph0):C (N15_180 ph15):N #endif SED_ON d30 H1_DEC_OFF TAUF SED_OFF d13 p18:gp4 ;1.25m, 30G/cm d16 (p2 ph16):N d6 (p1 ph0):H d13 p15:gp0 ;500u, 8G/cm TAUG (N15_180 ph0):N (H1_180 ph0):H d13 p15:gp0 ;500u, 8G/cm TAUG d7 (p2 ph1):N (p1 ph1):H d13 p15:gp0 ;500u, 8G/cm TAUG (C_H_N H1_180 ph0):H (N15_180 ph0):N d13 p15:gp0 ;500u, 8G/cm TAUG (p1 ph0):H TAUH (H1_180 ph0):H d13 p19:gp5*EA*-1 ;125u, -+30G/cm d18 pl12:N go=2 ph31 cpd2:N #ifdef ONE_D d11 do:N wr #0 #else d11 do:N wr #0 if #0 zd #endif #ifdef N15_EVOL 3m igrad EA d14 ip16 d14 ip16 lo to 3 times 2 d14 dd10 d14 id30 d14 ip14 d14 ip14 d14 ip31 d14 ip31 lo to 4 times l4 d14 rd10 d14 rd30 #endif #ifdef C13_EVOL d14 ip11 lo to 5 times 2 d14 id0 d14 dd20 d14 ip31 d14 ip31 lo to 6 times l6 #endif 20u LOCKH_OFF exit ph0=0 ph1=1 ph2=2 ph3=3 ph11=0 ph12=0 2 ph13=0 2 ph14=0 ph15=0 0 2 2 ph16=0 ph31=0 2