;hnca_trosy_H_ydw_v2 ;avance-version ;3D TROSY-HNCA for non-deuterated samples or proteins without deuteration at Ca ; F1(H) -> F3(N) -> F2(C=O,t1) -> F3(N,t2) -> F1(H,t3) ;phase sensitive (t1) ;phase sensitive using Echo/Antiecho (t2) ;using constant time in t2 ;M. Salzmann, K. Pervushin, G. Wider, H. Senn & K. Wuethrich, Proc. ; Natl. Acad. Sci. USA 95, 13585-13590 (1998) ;Water suppression: ; Using GS mode, adjust sp1 to minimize water signal ; Change gpz2 slightly to minimize water signal ;$OWNER=fan #include #include #include "p2=2*p1" "p22=2*p21" "d0=(in0-4*p3/3.1416-p22)/2" "d10=3u" "d11=30m" "d12=20u" "d13=5u" "d23=12.4m" "d21=1.34m" "d22=d23-d21-p2-p1-p1-6u" "d26=2.3m" "in30=in10" "d30=d23-p1" "DELTA=d1-2*d11-34m" "DELTA1=d26-p16-d16" "DELTA2=d23-d10-p14-p19-d16" "DELTA3=d26-p16-d16-p3-3u" "DELTA4=p22+d0*2+3u-6u-p14" "CEN_HN1=(p21-p1)/2" "CEN_HN2=(p22-p2)/2" "CEN_HN3=(p22-p2-p1-p1-6u)/2" "CEN_CN2=(p22-p14)/2" "l3=td1/2" "l13=td2/2" aqseq 321 1 d11 ze 2 d11 6m 3 18m 4 30m 5 9m 6 1m DELTA pl0:f1 pl5:f2 pl3:f3 50u UNBLKGRAD (p11:sp1 ph3:r):f1 4u 4u pl1:f1 (p1 ph1):f1 p16:gp1 d16 DELTA1 (CEN_HN2 p2 ph1):f1 (p22 ph1):f3 DELTA1 p16:gp1 d16 (p1 ph3):f1 p25:gp2 d16 ; N-->Ca (p21 ph9:r):f3 d21 (p1 ph2 3u p2 ph1 3u p1 ph2):f1 d22 (CEN_CN2 p14 ph1):f2 (p22 ph1):f3 ;13C 180 pulse center at 55ppm, p14=sqrt(3)/2/Delta d23 (p21 ph2):f3 p16:gp3 d16 pl2:f2 ; t1 period (p3 ph5):f2 ;13C 90 pulse at 55ppm, p3=sqrt(15)/4/Delta d0 pl0:f2 (CEN_HN3 p1 ph2 3u p2 ph1 3u p1 ph2):f1 (CEN_CN2 p14:sp5 ph10):f2 (p22 ph1):f3 ;sp5 center at 176ppm d0 pl2:f2 ; 3u pl5:f2 ; (p2 ph1):f1 (p14 ph1):f2 ; 3u pl2:f2 (p3 ph1):f2 (p1 ph2 3u p2 ph1 3u p1 ph2):f1 p18:gp4 d16 (p21 ph6):f3 d30 pl5:f2 (CEN_CN2 p14 ph1):f2 (p22 ph8):f3 p19:gp5*EA d16 DELTA2 pl0:f2 (p14:sp5 ph1):f2 d10 (CEN_HN1 p1 ph1) (p21 ph7):f3 3u pl2:f2 (p3 ph1):f2 p16:gp7 d16 DELTA3 (CEN_HN2 p2 ph1) (p22 ph1):f3 p16:gp7 d16 DELTA1 (CEN_HN1 p1 ph2) (p21 ph2):f3 p16:gp6 d16 DELTA1 (CEN_HN2 p2 ph1) (p22 ph1):f3 p16:gp6 d16 DELTA1 (CEN_HN1 p1 ph1) (p21 ph1):f3 p20:gp9 d16 4u (p2 ph1):f1 p20:gp8 d16 CEN_HN1 4u BLKGRAD go=2 ph31 d11 wr #0 if #0 zd 6m ip7*2 igrad EA lo to 3 times 2 6m id10 ip6*2 6m dd30 6m ip31*2 lo to 4 times l13 15m rd10 ip5 15m rd30 lo to 5 times 2 3m id0 3m 3m ip31*2 lo to 6 times l3 exit ph1=0 ph2=1 ph3=3 ph5=0 0 2 2 ph6=1 3 ph7=0 ph8=0 0 0 0 2 2 2 2 ph9=0 ph10=0 0 0 0 2 2 2 2 ph11=2 ph31=0 2 2 0 ;pl0 : 120dB ;Channel 1 f1 ;PL1: power level for hard high power pulse ;p1: 90 degree pulse @PL1 ;p2: p2=2*p1, 180 degree pulse @PL1 ;sp1: power level for shaped pulse ;p11: 90 degree pulse @sp1 ;spnam1: eburp2 shape (~5ms) center at water ;spoffs1: 0Hz, offset for sp1 ;O1p --> 4.7ppm on water resonance ;Channel 2 f2 ;PL2: power level for hard 90 degree pulse ;p3: 90 degree pulse @pl2, p3=sqrt(15)/(4*delta), ; delta=(176-55)*spectrometerfrequency ;PL5: power level for hard 180 degree pulse ;p14: 180 degree pulse @PL5 or @sp5, p14=sqrt(3)/(4*delta), ; delta=(176-55)*spectrometerfrequency ;sp5: power level for shaped pulse, sp5=PL5 ;spnam5: shaped hard, hard.100; center at 176 ppm ;spoffs5: offset for sp5, (176-55)ppm, 24345Hz @800MHz, 18259Hz @600MHz ;O2p=53ppm, corresponding to ~55.7ppm relative to TSP ;Channel 3 f3 ;PL3: power level for high power pulse ;p21: 90 degree pulse @PL3 ;p22: p22=2*p21 ;O3p=119ppm ;Gradients ;gpnam1: SINE.100 ;gpnam2: SINE.100 ;gpnam3: SINE.100 ;gpnam4: SINE.100 ;gpnam5: rectangular ;gpnam6: SINE.100 ;gpnam7: SINE.100 ;gpnam8: rectangular ;gpnam9: rectangular ;p16: gradient pulse @gpz1, gpz3, gpz6 and gpz7 [1400 usec] ;gpz1: 50%; gpz3: 30%; gpz6: 30%; gpz7:-30% ;p18: gradient pulse @gpz4 [1200 usec] ;gpz4: 35% ;p19: gradient pulse @gpz5 [1250 usec] ;gpz5: -80% ;p20: gradient pulse @gpz8 and gpz9 [63.4 usec] ;gpz8: 80%; gpz9: -80% ;p25: gradient pulse @gpz2 [1000 usec] ;gpz2: ~30% ;Other parameters ;in0: 1/(2*SW(C)), SW(C)=~20 ppm (in the TD1 dimension) ;d0: incremented delay (in Ca) [-90/180 phase correction] ;MAKE SURE d0>0, use ased to check ;nd0: 2 ;in10: 1/(2*SW(N)) ;nd10: 2 ;NS: 4*n ;DS: 16