;hncacb.ref, av700 version 09/02/2002 ;history ;written on 10/23/91 ;used for sg hncoct/8 10/23/91 ;interchanged C13 and N15 6/12/92 ;used for gv ;put in NH2 purge pulse ph22 7/17/92 ;put additional +/- on last nitrogen 90 pulse 7/17/92 ;used for jacob's calcineurin 7/17/92 ;changed to hnco_fb.sg 9/1/94 included flip-back ;used on NEF hncoct/10 SG 9/1/94 ;changed to hnca_fb.sg 9/2/94 ;used on NEF hnca/46 SG 12/94 ;changed to hncacb_fb.sg 1/26/95 ;changed to hncacb_fb.kf for the DMX500 1/24/96 (by Daron Freedberg and Irina Kustanovich) ;adopted to dmx750 by yxw 12/16/96 ;adapted to drx600 by aph 12/28/01 ;adapted to drx700 by aph 9/02/02 #include "Grad.incl" ;Adjust these as appropriate by commenting in/out ;CALPHA turns on evolution of 13c ;NITROGEN turns on evolution of 13c #define CALPHA ;#define NITROGEN ;Channel assignments; adjust according to your settings ;in "edasp" #define H1 f1 #define C13 f2 #define N15 f3 ;Proton settings - Set carrier to H2O ;p1 - square 90 1H @ pl1, for excitation, typically p1 = 7.9u @ pl1 = -2dB ;p2 - square 90 1H @ pl1, for watergate, typically p2 = 1000u @ pl2 = +39.6dB ;p3 - shaped 90 1H @ sp0, for water flipback, typically p3 = 2m @ sp0 = +39.2 dB ; "spnam0 seduce1.jc", "spoffs0 0" ;pcpd1 - square 90 1H @ pl30, for excitation, typically pcpd1 = 50u @ pl30 = +13.2dB ; "cpdprg1 dipsi2" ;Carbon settings - Set carrier to 46.0 ppm ;p11 - square 90 13C @ pl11, for excitation, p11 must equal 42.3u, typically pl11 = +5.0 ;p12 - square 90 13C @ pl12, for inversion, p12 must equal 18.9u, typically pl12 = -2.1 ;p13 - shaped 180 13C @ sp1, for CO inversion, typically p13 = 200u @ sp1 = +7.5 dB ; "spnam1 sinc1.100", "spoffs1 22886" ;Nitrogen settings - Set carrier to amide center (ca. 118 ppm) ;p7 - square 90 15N @ pl7, for excitation, typically p7 = 45u @ pl1 = -3.8dB ;pcpd3 - square 90 15N @ pl31, for cpd, typically pcpd3 = 200u @ pl31 = +8.7 ; "cpdprg3 waltz16" ;15N indirect dimension ;in21=in22=in23=1/(4sw), nitrogen spectral window, typically in21=146.8u ;l4, nitrogen t1 points, typically l4 = 45 ;zero order phase = 0.0 deg ;first order phase = 0.0 deg ;reverse = no ;13C indirect dimension ;in0=1/(2sw), carbon spectral window, typically in0=42.4u ;l3, carbon t1 points, typically l3 = 89 ;zero order phase = -90.0 deg ;first order phase = 180.0 deg ;reverse = no ;GRADIENT SETTINGS ;Note1 - 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.32", "gpx0 20", "gpy0 0", "gpz0 -40" ; ;grad# gpnam# gpx# gpy# gpz# ;0 sine.32 20 0 50 ;These are the gradient lengths; These MUST be entered manually ;at the XWIN-NMR command line ;p20=1m at gp0=+50% ;p19=1m at gp1=+50% ;p18=500u at gp2=+50% ;p17=750u at gp3=+50% ;p16=1.5m at gp4=+50% ;p15=400u at gp5=+50% ;DELAYS THAT MAY REQUIRE SLIGHT ADJUSTMENT "d1=1.0s" ;recycle time "d7=4u" ;preacquisition delay, adjust for zero order phase = 0.0 ;DELAYS - DO NOT REQUIRE ADJUSTMENT! ;#define TEST_1D #define ON #ifdef CALPHA "d0=in0 - p11*0.637" ;180/-360 (bruker), -180/+360 (nmrPipe) #else "d0=2u" #endif "d2=p1" "d3=p12*2" "d4=2.25m" ;1/4J for NH coupling "d5=p12*2" "d11=50m" "d12=7m" ;universal dd,ip delay "d22=6.81m" ;For 15N Evolution "d21=d22-5.4m" ;For 15N Evolution "d23=d22" ;For 15N Evolution "d26=p7-p1" "d28=81u - p7" ;****************END DECLARATION ******************** ze 10u 2 d11*0.5 do:N15 d11*0.5 BLKGRAD #ifdef CALPHA 21 d12*4 3 d12*2 4 d12 #endif #ifdef NITROGEN d12 5 d12*4 6 10u #endif 10u d1 pl7:N15 1m UNBLKGRAD (p7 ph10):N15 p20:gp0 50u pl1:H1 50u pl7:N15 #ifdef ON ;******* start 90-degree ******* (p1 ph0) d4 (d26 p1*2 ph1) (p7*2 ph2):N15 d4 (p1 ph2) 2u 50u pl2:H1 (p2 ph24):H1 2u p19:gp1 2m pl1:H1 (p7 ph3):N15 ;******* n15 evolution ******* 5.4m (p1 ph26) ;adjust ph26 to be along y for relative phases of hard 90 and dipsi 4u pl30:H1 ;set power level for DIPSI d21 cpds1:H1 ;start DIPSI-2 on 1H channel (p13:sp1 ph10):C13 ;shaped 180 degree sinc1.0 on carbonyls 2u d22 pl12:C13 d3 2u (p7*2 ph5):N15 2u (p12*2 ph6):C13 2u d23 (p13:sp1 ph10):C13 d23 d2 6u pl11:C13 (p7 ph10):N15 ;******* apply z-filter ******* 4u do:H1 4u pl1:H1 (p1 ph27) 4u p18:gp2 800u (p1 ph26) 2u 4u pl30:H1 30u cpds1:H1 ;restart 1H DIPSI ;******* end z-filter ******* (p11 ph4):C13 ;******* inept to c-alpha/beta ******* 1m 2.5m pl12:C13 (p12*2 ph22):C13 2.5m 1m pl11:C13 ;******* c-alpha/beta evolution ******* (p11 ph7):C13 d0 d0 (p11 ph8):C13 ;******* inept from c-alpha/beta ******* 1m 2.5m pl12:C13 (p12*2 ph9):C13 2.5m 1m pl11:C13 (p11 ph20):C13 ;******* apply z-filter ******* 4u do:H1 4u pl1:H1 (p1 ph27) 4u p17:gp3 1.2m (p1 ph26) 2u 4u pl30:H1 30u cpds1:H1 ;******* end z-filter ******* (p7 ph10):N15 4u 4u pl12:C13 d2 10m d5 2u (p7*2 ph10):N15 2u (p12*2 ph20):C13 4.6m 4u do:H1 4u pl1:H1 (p1 ph27) 5.4m (p7 ph0):N15 5u p16:gp4 3m (p3:sp0 ph25):H1 2u 50u pl1:H1 (p1 ph0) 5u p15:gp5 950u pl2:H1 (p2 ph24):H1 2u 5u pl1:H1 (p1*2 ph0) 2u 5u pl2:H1 (p7*2 ph10):N15 (p2 ph24):H1 3u (2u ph0) p15:gp6 950u pl31:N15 d7 #endif #ifndef TEST_1D go=2 ph31 cpd3:N15 1m BLKGRAD d11 do:N15 wr #0 if #0 zd #ifdef CALPHA d12 ip4 d12 ip4 d12 ip4 d12 ip7 lo to 3 times 2 d12 id0 d12 ip31*2 lo to 4 times l3 d12 rd0 #endif #ifdef NITROGEN d12 ip3 lo to 5 times 2 d12 id21 d12 id22 d12 dd23 d12 ip31*2 lo to 6 times l4 #endif #else ; 950u 10u pl1:H1 p1 ph21 (2u ph0) go=2 ph31 d11 wr #0 #endif 5u do:N15 10u exit ph0=0 ph1=1 ph2=1 ph3=0 ph4=0 2 ph5=0 0 0 0 1 1 1 1 ph6=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 ph7=1 1 3 3 ph8=1 ph9=1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 ph10=0 ph11=(6)0 ph12=(6)5 ph13=(6)4 ph14=(6)3 ph15=(6)2 ph16=(6)1 ph20=0 ph21=0 ph22=0 ph24= 2 ;adjusted -x at hl2 set phcor24 ph25= 2 ;adjusted -x at sp0 ph26= 1 ;(3600) 1014 ;adjusted y for pl1 relativ to pl3 ph27= 3 ; =(3600) 2814 ;adjusted -y for hl1 relativ to hl3 ph31=0 2 0 2 2 0 2 0 2 0 2 0 0 2 0 2