;;Dimensionality: 5 ;;Experiment: BAONH ;;Phaseoverlap: 00000 ;;trosy: 0 ;;DimProjections: 2 ;;Hfreq: BF1 ;;Cfreq: BF2 ;;Nfreq: BF3 ;;Carrier1: CNST 31 ;;Carrier2: CNST 31 ;;Carrier3: CNST 29 ;;Carrier4: O3P ;;Carrier5: O1P ;;----------------end header------------------ ;USERA5: 5D BAONH st ;Conventional name: 5D-APSY-CBCACONH ;Sebastian Hiller, 2004, optimized 6/2005 ;modified for manageapsy: Fred Damberger ETH ZH 2007 ;**************************************************************** ;* * ;* The following one-letter codes for the nuclei are used: * ;* * ;* CB = B v w1 * ;* CA = A w w2 * ;* CO = O x w3 * ;* N = N y w4 * ;* H = H z w5 * ;* * ;* * ;* The angles are defined as: * ;* * ;* alpha between y and x * ;* beta between xy and w * ;* gamma between wxy and v * ;* * ;* * ;**************************************************************** ;pl1 : power for 1H hard pulse ;pl2 : power for 13C hard pulse ;pl3 : power for 15N hard pulse ;pl9 : power for H2O presat ;pl11 : power for 1H DIPSI2 decoupling ;pl13 : power for 15N WALTZ decoupling ;p1 : 90 degree hard pulse 1H ;p2 : 180 degree 1H ;p3 : 90 degree hard pulse 13C ;p4 : 180 degree 13C ;p5 : 90 degree hard pulse 15N ;p6 : 180 degree 15N ;p13 : 180 degree 13Cab (null@CO) ;p14 : 90 degree 13Cab (null@CO) ;p18 : 180 degree [80u] gauss128_5 ;p9 : 180 degree [220u] iburp1_128 ;p25 : purge pulse 1m ;pcpd1 : 90 degree cpd-pulse 1H (DIPSI2, 80us) ;pcpd3 : 90 degree cpd-pulse 15N (WALTZ, 160us) ;cpdprg1 : dipsi2 ;cpdprg3 : waltz16 ;p21 : 600u ;p22 : 800u ;p23 : 1000u ;gpz1 : 25 % ;gpz2 : 30 % ;gpz3 : 35 % ;gpz4 : 40 % ;gpz5 : 45 % ;gpz6 : 50 % ;gpz7 : 55 % ;gpz8 : 60 % ;gpz9 : 65 % ;gpz10 : 70 % ;gpz11 : 75 % ;gpz12 : 80 % ;d1 : relaxation delay ;d5 : 3-9-19-delay. 200/130u at 500/750MHz ;cnst1: alpha ;cnst2: beta ;cnst3: gamma ;cnst41: SW (CB) [Hz] ;cnst42: SW (CA) [Hz] ;cnst43: SW (CO) [Hz] ;cnst44: SW (N) [Hz] "cnst41=cnst41" "cnst42=cnst42" "cnst43=cnst43" "cnst44=cnst44" ;cnst7: 1H carrier in HA ;cnst8: 1H carrier in amide region ;cnst29: CO chemical shift (offset, 174 ppm) ;cnst30: Ca chemical shift (offset, 53 ppm) ;cnst31: Cab (or ali) chemical shift (offset, 43ppm) ;cnst51: set to CNST31 ;cnst52: set to CNST31 ;cnst53: set to CNST29 ;cnst54: set to O3P ;cnst55: set to O1P "cnst51=cnst51" "cnst52=cnst52" "cnst53=cnst53" "cnst54=cnst54" "cnst55=cnst55" ;sp9 : 180 deg, p9, iburp1_128 ;sp13 : 180 deg, p13, BLOCK.32 ;sp14 : 90 deg, p14, BLOCK.32 ;sp16 : 90 deg, p16, BLOCK.32 ;sp17 : 180 deg, p17, BLOCK.32 ;sp18 : 180 deg, p18, gauss128_5 ;spnam7 : BLOCK.32 ;spnam8 : BLOCK.32 ;spnam9 : iburp1_128 ;spnam13 : BLOCK.32 ;spnam14 : BLOCK.32 ;spnam16 : BLOCK.32 ;spnam17 : BLOCK.32 ;spnam18 : gauss128_5 ;spnam21 : gauss128_5 #define GRADIENT1 10u p22:gp3 190u #define GRADIENT2 10u p22:gp7 190u #define GRADIENT3 10u p22:gp1 190u #define GRADIENT4 10u p22:gp5 190u #define GRADIENT5 10u p22:gp10 190u ;----------------------offsets "p13=866000/(bf2*(cnst29-cnst30))" "p14=968000/(bf2*(cnst29-cnst30))" "p16=968000/(bf2*(cnst29-cnst30))" "p17=866000/(bf2*(cnst29-cnst30))" "spoff9=bf2*((cnst30-cnst29)/1000000)" "spoff13=0.0" "spoff14=0.0" "spoff16=0.0" "spoff17=0.0" "spoff18=bf2*((cnst29-cnst31)/1000000)" "in11=abs(in1*sin(0.0174533*cnst3))/2" ; CB "in12=abs(in1*sin(0.0174533*cnst2)*cos(0.0174533*cnst3))/2" ;CA "in13=abs(in1*sin(0.0174533*cnst1)*cos(0.0174533*cnst2)*cos(0.0174533*cnst3))/2" ;CO "in14=abs(in1*cos(0.0174533*cnst1)*cos(0.0174533*cnst2)*cos(0.0174533*cnst3))/2" ;N "l3=td1/2" "p2=2*p1" "p6=2*p5" "d0 = 1.0m" ; dummy "in0= in1" ; dummy ;------------------------------basic delays "d2 = 2.7m" ; 1/2 H-N "d3 = 1.8m" ; 1Hb-13Cb inept delay "d4 = 5.4m" ; 1/2J H-N "d6 = 2.3m" ; delay for 1H in-phase "d13 = 4.75m" ; 13CO-13CA inept delay "d9 =11.4m" ; 13CO-15N inept delay "d8=11.0m" ; 1/2 constant-time 13CO-15N "d19= 3.3m" ; 1/2 constant-time 13C "d10= 6.75m" ;----------------------fixed delays define delay IN_HN "IN_HN=d2-(p22+200u+p1*2.3846+d5*2.5)" define delay LAMBDA "LAMBDA=d5/2-p5" ;----------------------------indirect dimensions "d17=d3-p22-200u" ;----------------------B "d11=d19" "d21=10u" "d22=d19" "in22=in11" ;----------------------A "d12=d13" "d24=d10-d13" "d25=d10-5u" "in25=in12" ;----------------------O "d27=d9-d13-p6" "d26=10u" "in9=(d9-20u)/l3" "in26=0u" ;----------------------N "d31=d8" "d28=10u" "d16=d8-p17-10u-d4" "d14=d8-p17-10u" "d29=d8-p9-p17-20u" "in31=(d31-20u)/l3" "in16=in14" "in28=0u" ;-------------------- #include 1 10u ze #ifdef A2 3m ip31*2 #endif #ifdef V 3m ip11 #endif #ifdef W 3m ip12 #endif #ifdef X 3m ip13 #endif #ifdef Y 3m dp14 #endif goto 44 10u d0 10u id0 10u id22 10u id25 44 1m ;------------------------------------Switch constant-time for t4(15N) [dim y] if "in31 < in14" goto 5 10u "in31=in14" 10u goto 6 5 1m 10u "in28=in14-in31" 10u 6 1m ;------------------------------------Switch constant-time for t3(13CO) [dim x] if "in9 < in13" goto 7 10u "in9=in13" 10u goto 8 7 1m 10u "in26=in13-in9" 10u 8 1m ;-----------------------------------Start 2 1m 3 1m 4 1m 10u do:f3 10u LOCKH_OFF 10u pl9:f1 d1 cw:f1 10u do:f1 100u fq=cnst31 (bf ppm):f2 20u pl1:f1 20u pl2:f2 20u pl13:f3 20u LOCKH_ON ;-----------------------------------------INEPT 1H13CB (p1 ph20):f1 GRADIENT1 d17 (refalign (p13:sp13 ph20):f2 center (p2 ph20):f1) GRADIENT1 d17 (p1 ph21):f1 ;-----------------------------------------z filter GRADIENT2 10u cpds3:f3 10u pl11:f1 ;-----------------------------------------13CB evolution (p14:sp14 ph11):f2 5u (refalign (d11 p18:sp18 ph20 d21):f2 lalign (d6 5u cpds1):f1) (p13:sp13 ph20):f2 d22 (p18:sp18 ph20):f2 15u ;-----------------------------------------INEPT 13Ca13CO & 13CA evolution (p14:sp14 ph7):f2 d12 (p18:sp18 ph20):f2 d24 (p13:sp13 ph20):f2 d25 (p18:sp18 ph20):f2 5u (p14:sp14 ph12):f2 ;-----------------------------------------zzz 10u do:f3 GRADIENT5 10u fq=cnst29 (bf ppm):f2 10u 10u pl2:f2 10u pl3:f3 ;-----------------------------------------INEPT 13CO15N & 13CO evolution (p16:sp16 ph8):f2 d13 (p9:sp9 ph20):f2 d27 (p6 ph20):f3 d26 (p17:sp17 ph20):f2 d9 (p9:sp9 ph20):f2 10u (p16:sp16 ph13):f2 ;-----------------------------------------z filter GRADIENT3 ;-----------------------------------------15N evolution (p5 ph14):f3 d31 (p6 ph20):f3 d28 (p17:sp17 ph20):f2 (d16 5u do 5u pl1):f1 (d29 p9:sp9 ph20):f2 (d14):f3 (p5 ph20):f3 ;-----------------------------------------INEPT 15N1H (p1 ph20):f1 GRADIENT4 IN_HN (p1*0.2308 ph21 d5 p1*0.6923 ph21 d5 p1*1.4615 ph21):f1 (LAMBDA p6 ph20 LAMBDA):f3 (p1*1.4615 ph23 d5 p1*0.6923 ph23 d5 p1*0.2308 ph23):f1 GRADIENT4 IN_HN pl13:f3 ;-----------------------------------------acquisition go=2 ph31 cpd3:f3 1m do:f3 wr #0 if #0 zd #ifdef Pv 30u ip11 #endif #ifdef Pw 30u ip12 #endif #ifdef Px 30u ip13 #endif #ifdef Py 30u dp14 #endif lo to 3 times 2 1m id11 1m dd22 1m id12 1m dd25 1m id13 1m id26 1m dd9 1m id14 1m id16 1m id28 1m dd31 lo to 4 times l3 10u do:f1 10u do:f2 10u do:f3 10u LOCKH_OFF exit ;-------------------phases for phasecycle ph8= 0 2 ph7= 0 0 2 2 ph31= 2 0 ;-------------------fixed phases ph20=0 ph21=1 ph22=2 ph23=3 ;-------------------phases for States-TPPI ph11=0 ph12=0 ph13=0 ph14=0