;HNCA[HA]-E.COSY for the Bruker AMX-600. ;3J(HnHa) from E.COSY splitting. ;Andy C. Wang and Ad Bax, March 1995 ;JACS submitted, October, 1995 ;BMRB Pulse Sequence Accession Number: 33 ;_______________________________________________________________________ #include "bits.sg" #define CARBON #define NITROGEN #undef COADD_2D #define CARBON_LOOP 24 #define NITROGEN_LOOP 32 #define ITERATIONS 9 ;_______________________________________________________________________ ;PROTON pulses: ;carrier at 8.36ppm ;p1 = 90 at hl1=3dB ;p2 = 720us proton 90 @ tp0=38.5dB and tpname0=sinc1.0 ;p4 = 890us proton 90 @ tp1=29.5dB and tpname1=q3_128.acw ;p30 = 50us 90 at hl3 = 17dB for Dipsi-2 decoupling ;NITROGEN pulses: ;p3=nitrogen 90 at dbhi ;p31=nitrogen 90 at dbl0 ;CARBON pulses: ;carrier at 58 ppm ;p5=53.9u Ca 90 @ dl0=29 dB for null at 177 ppm ;p6=24.1u Ca 90 @ dl1=21 dB for null at 177 ppm ;p7=97.4u C' 90-deg offset sinc used as 180-deg @ dp0=30 dB ;p9=150u C' 90-deg offset sinc used as 180-deg @ dp2=33 dB ;_______________________________________________________________________ ;Nitrogen constant time: d18=6.81m d10=d18 d20=d18 ;in10=in19=in20 ;in10=in20=in19=1/(4*sw) ;CARBON constant time: ;sw=1/(2*in0) d0=in0-p9-4u-p5*0.636 ;_______________________________________________________________________ ;DELAYS: d3=2.25m ;d7 ca 100u = dw d8=aq d11=50m d12=8m d23=p3-p6 ;p3 > p6 d24=p7-p1 d26=p3-p1 d27=p9-p3 d28=p4-p3 ;_______________________________________________________________________ ;f1 = hydrogen frequency on water ;f2 = Ca frequency 58 ppm ;f3 = nitrogen frequency ;_______________________________________________________________________ 100u CLEAR_BITS ;switch 2H decoupler off, switch everything off 100u LOCK_ON ;switch lock on ze 2 d11 dbo d12 3 d12*3 4 d12*2 5 d12*4 6 d12*6 7 10u dbo 10u do 10u to 100u LOCK_ON ;switch lock on d1 dlo 20m hl1 20u dbhi ;******* Start 90-degree ******************************** (p1 ph0):f1 d3 (d26 p1*2 ph10):f1 (p3*2 ph0):f3 d3 (p1 ph1):f1 50u LOCK_OFF GRAD(10, POSITIV, 40) 3m (p3 ph3):f3 ;******* N15 evolution ******************************** d10 dl6 4u (p7*2 ph10):dp0 (d24 p1*2 ph0):f1 d10 dl1 10u (d23 p6*2 ph0):f2 (p3*2 ph5):f3 10u d20 dl6 (p7*2 ph10):dp0 (d24 p1*2 ph0):f1 4u d20 dl0 (p3 ph1):f3 GRAD(20, POSITIV, 30) 2.25m (p5 ph4):f2 ;******* C-alpha evolution ******************************** 4u d0 dl6 ;t1/2 (d27 p3*2 ph10):f3 (p9*2 ph20):dp2 d0 4u dl0 ;******* back to N ******************************** (p5 ph6):f2 4u (p3 ph10):f3 11m 4u dl1 (d23 p6*2 ph0):f2 (p3*2 ph10):f3 4u 11m tlo ;******* back to Hn ******************************** (p3 ph1):f3 GRAD(30, POSITIV, 20) 1.5m (p2 ph8):tp0 GRAD(40, POSITIV, 4) d4 (p4*2 ph7):tp1 (d28 p3*2 ph10):f3 GRAD(50, POSITIV, 4) (2u ph0) d7 dblo 2u dbl0 p24:e 3u adc d8 cpdb 10u do 10u to 10u dbo rcyc=2 ph31 d11 wr #0 if #0 zd ;_______________________________________________________________________ #ifdef CARBON d12 ip4 lo to 3 times 2 d12 id0 d12 ip31 d12 ip31 lo to 4 times CARBON_LOOP #endif ;_______________________________________________________________________ #ifdef NITROGEN d12 ip3 d12 rd0 lo to 5 times 2 d12 id10 d12 dd20 d12 ip31 d12 ip31 lo to 6 times NITROGEN_LOOP #endif ;______________________________________________________Co-add 2D spectra #ifdef COADD_2D d12 rf #0 d12 rp4 d12 rp31 d12 rd0 d12 CLEAR_BITS d12 ze lo to 7 times ITERATIONS #endif ;_______________________________________________________________________ 100u do 100u dbo 100u to 100u LOCK_ON ;switch lock on 100u CLEAR_BITS ;switch 2H decoupler off, switch everything off exit ;_______________________________________________________________________ ph0=0 ph1=1 ph3=0 ph4=0 ph5=0 1 2 3 ph6=0 0 2 2 ph7=(360) 327 ph8=0 0 0 0 2 2 2 2 ph10=0 ph20=0 ph31=0 2 2 0 2 0 0 2 ;_______________________________________________________________________