;noesy_nc4dshare_ydw_v6 ;avance-version ;4D 13C/15N-edited NOESY (HMQC-NOESY-HSQC), used for proteins larger than 25 kDa ;F1 1H (constant-time acq mode, maximum aq time 6.4ms), F2 13C/15N (both aliphatic and aromatic), ;F3 13C/15N, and F4 1H ;without gradient enhancement ;water suppresion with water gate; ;Written by Daiwen Yang in Mar. 2006 ; modified in Apr. 2009: Replacing the water gate (90s18090s) scheme to the W5 scheme ; Changing all 1H 180 pulses to composite 180 pulses ; Acquiring indirect 1H first ;Water Suppression: ; Using GS mode, 1). adjust p0 to minimize water signal ; 2). adjust sp1 to minimize water ; 3). adjust phcor10 ;with proper values of p0, sp1 and phcor10, the water signal intensity can be comparable to ;CH3 signals even for a cryoprobe on 800MHz without applying any solvent filter processing ; set d18 (delay used in the W5 scheme), 170us @800MHz, 220us@600MHz ;$OWNER=fan #include #include #include "p2=p1*2" "p4=p3*2" "d0=(in0-p5*4/3.1416-p2-p1-p1-3u-3u)/2" "d10=(in10-p5*4/3.1416-p1-3u-p2-3u-p1-32u)/2" ;modified to compensate for long p1 "d19=(in19-p3*4/3.1416-2*p5+p5*4/3.1416-40u)/2" "d20=3u" "DELTA1=2*p5-p5*4/3.1416" "d22=1.8m" "d23=2.2m" "d4=d22-p14/2" "d5=d23-d22-p14/2" "d6=d23-p19-d16-4.5*d18-2.888*p29-0.087*p29/2-0.087*p0/2-p1*2/3.1416" "d21=d23-p19-d16-4.5*d18-2.888*p29-0.087*p29/2-0.087*p0/2-4u" "d7=(d23-d22)*2" "d24=1.7m" "d25=1.91m" "d26=d24-p16-d16-p12/2-d20" "d27=d25-d24-p12/2" "d28=d24-p16-d16-d20-DELTA1-p4-p12/2" "d29=d25-p16-d16" "d11=30m" "d13=3u" "CEN_HN2=(p4-p2)/2" "d8=d9-p18-d16-p3-p5" "DELTA=d1-d11-5m-10m-12m-18m-10m-10m-10m" "in29=in20" "l3=(td1/2)" "l13=(td2/2)" 1 ze d11 pl12:f2 pl13:f3 2 d11 do:f3 do:f2 5m 3 10m 4 12m 5 18m 6 10m 7 10m 8 10m do:f3 DELTA pl3:f3 pl2:f2 pl1:f1 50u UNBLKGRAD 10m (p5 ph1):f2 (p3 ph1):f3 p20:gp6 d16 pl0:f2 (p1 ph8):f1 p16:gp2 ;p16 200u d16 ;d16 100u d20 d26 ; 1.7m-p16-d16-d20-p12/2 (p12:sp2 ph1):f2 d27 ; 2.0m-1.7m-p12/2 (center (p1 ph1 3u p2 ph2 3u p1 ph1):f1 (p4 ph1):f3) p16:gp3 d16 pl2:f2 d29 ; 2.0m-p16-d16 (p3 ph5):f3 (p5 ph6):f2 d0 ;t1 period for 13C and 15N (p1 ph1 3u p2 ph2 3u p1 ph1):f1 d0 (p5 ph1):f2 (p4 ph1):f3 DELTA1 ;2*p5-p5*4/3.1416 (p3 ph1):f3 p16:gp7 d16 pl0:f2 d20 d28 ; 1.7m-p16-d16-d20-DELTA1-p4-p12/2 (p12:sp2 ph1):f2 d27 (center (p1 ph1 3u p2 ph2 3u p1 ph1):f1 (p4 ph1):f3) p16:gp8 d16 pl2:f2 d29 ; 2.0m-p16-d16-3u (p1 ph1:r):f1 ;+/-45 deg, water supppression d8 ;d8=d9-p18-d16-p3-p5 (p5 ph1):f2 (p3 ph1):f3 p18:gp4 d16 pl0:f2 (p1 ph1):f1 d4 ;1.8m-p14/2 (p14:sp3 ph1):f2 d5 ;2.2m-1.8m-p14/2 (center (p2 ph2):f1 (p4 ph1):f3) d23 ;2.2m (p1 ph2):f1 3u pl11:f1 (p11:sp1 ph11:r):f1 ;optimize water suppression via phcor11 3u p17:gp1 d16 pl1:f1 (p3 ph9):f3 5u pl22:f2 5u cpds4:f2 d19 5u do:f2 5u pl2:f2 (p5 ph3):f2 d10*0.5 gron0 8u groff d10*0.5 gron0*-1 8u groff (p1 ph2 3u p2 ph1 3u p1 ph2):f1 d10*0.5 gron0*-1 8u groff d10*0.5 gron0 8u groff (p5 ph1):f2 5u pl22:f2 5u cpds4:f2 d19 5u do:f2 5u pl2:f2 (p3 ph1):f3 p15:gp9 d16 pl0:f2 3u pl11:f1 (p11:sp1 ph10:r):f1 3u 3u pl1:f1 (p1 ph1):f1 d6 p19:gp5 d16 pl18:f1 ; d6: 2.2m(d23)-p19-d16-4.5*d19-2.888*p28-0.087*p28/2-0.087*p0/2-p1*2/3.1416 (center (p29*0.087 ph12 d18 p29*0.206 ph12 d18 p29*0.413 ph12 d18 p29*0.778 ph12 d18 p29*1.491 ph12 d18 p29*1.491 ph13 d18 p29*0.778 ph13 d18 p29*0.413 ph13 d18 p29*0.206 ph13 d18 p0*0.087 ph13):f1 (p14:sp3 ph1 d7):f2 (p4 ph1):f3) p19:gp5 d16 d21 pl12:f2 pl13:f3 ;d21 2.2m(d23)-p19-d16-4.5*d19-2.888*p28-0.087*p28/2-0.087*p0/2-4u 4u BLKGRAD go=2 ph31 cpds2:f2 cpd3:f3 d11 do:f2 do:f3 wr #0 if #0 zd 5m ip8 lo to 3 times 2 3m id20 3m dd29 4m ip31*2 lo to 4 times l23 3m rd20 3m rd29 3m ip3 3m ip9 lo to 5 times 2 6m id10 6m id19 6m ip31*2 lo to 6 times l13 2m rd10 2m rd19 2m ip6 4m ip5 lo to 7 times 2 5m id0 5m ip31*2 lo to 8 times l3 exit ph1=0 ph2=1 ph3=0 0 2 2 ph5=0 2 ph6=0 2 ph8=0 ph9=0 0 2 2 ph10=0 ph11=2 ph12=0 ph13=2 ph31=0 2 2 0 ;Channel 1 1H ;o1p: 4.7ppm (water) ;p0: ~90 degree hard pulse@PL18 ;p1: 90 degree hard pulse @PL1 (as short as possible) ;p2: 180 degree hard pulse P2=2*P1 ;p18: 90 degree hard pulse@PL18 ;p11: 90 degree rectangle-shaped pulse @SP1, ~2ms; on-resonance ;Channel 2 13C ;o2p: 54ppm (in spectrometer parameter), corresponding to 56.7 ppm relative to TSP ;p5: 90 degree hard pulse @PL2 (as short as possible) ;PL12: adiabadic decoupling power level during 1H acq, ~4.2kHz @800MHz, 3.7kHz@600MHz, 3.5kHz@500MHz ;PL22: CO-decoupling power level during 15N acq, ~4.3kHz @800MHz, 3.3kHz @600MHz, 2.7kHz@500MHz ;p12: adibadic 180 pulse @sp2, 400us ;sp2: power level, rf=13.7kHz ;spnam2: CaWurst400_65k ;spoffs2: offset for sp2, 0Hz ;p14: adibadic 180 pulse @sp3, 600us ;sp3: power level, rf=10.3kHz ;spnam2 CaWurst600_55k ;spoffs3: offset for sp3, 0Hz ;cpdprg2: p5m4sp180 broad band C decoupling (pcpd2=1.2ms, @sp15) ;pcpd2=1.2ms ;sp15: decoupling power level, sp15=PL12, rf: ~4.2kHz @800MHz ;spnam15: Crp_1200_45k_20_800, smoothedChirp ;spoffs15: (76-56)ppm, 4024Hz @800MHz ;cpdprg4: mlevsp180_ydw carbonyl ;p28: 250 us @800 MHz, 330us@600MHz, 400us@500MHz ;sp13: CO-decoupling power level,sp13=PL22 ;spnam13 sed.250, seduce shape ;spoffs13: (176-56)ppm, 24144Hz @800MHz ;NOTE: sp13 and sp15 do not appear in paeameter list in ased. INPUT IN MANUALY!!!! ;Channel 3 15N ;o3p: 119ppm ;p3: 90 degree hard pulse @PL3 ;p4: 180 degree hard pulse, P4=2*P3 ;PCPD3: 90 degree pulse @PL12, 250us at 800 MHz; 300us at 600MHz and 500MHz ;CPDPRG3: GARP ;Gradients ;gpname1: SINE.100 ;gpname2: RECT.1 ;gpname3: RECT.1 ;gpname4: SINE.100 ;gpname5: SMSQ10.50 ;gpname6: SINE.100 ;gpname7: RECT.1 ;gpname8: RECT.1 ;gpname9: SINE.100 ;p15: gradient pulse @gp9, [1000us] ;gpz9: 20% ;p16: gradient pulse @gp2, gp3, gp7 and gp8 ;gpz2: 10% ;gpz3: 30% ;gpz7: 40% ;gpz8: 20% ;DO NOT CHANGE gpz2, gpz3, gpz7 and gpz8 ;p17: gradient pulse @gp1 ;gpz1: 15% ;p18: gradient pulse @gp4 ;gpz4: -5% ;p19: gradient pulse @gp5 ;gpz5: 50% ;p20: gradient pulse @gp6 ;gpz6: 10% ;Other Parameters ;in0: 1/SW(C1)/2=1/SW(N1) in the HMQC period; SW(C1)=22ppm ;in10: 1/SW(C2)/2 in the HSQC period; SW(C2)=22ppm ;in19: 1/SW(N2)/2-1/SW(C2)/2 in the HSQC period; SW(N2) similar to the HNCA or HSQC one, ; try to minimize it by folding a few peaks. ;in20: 1/SW(H)/4; SW(H) ~10ppm, covering all protons. ;in20 must be typed in manually, if it is different from the default one (31 us) ;in29: =in20 ;l3: td1/2 ;l13: td2/2 ;l23: 52 - 30, must be typed in manually. ;d18: delay used in the W5 scheme, 170us @800MHz, 220us@600MHz ;d9: NOE mixing time, ;ns: 4*n ;ds: 32