; DFhncocacbDwgnltp ; avance/CRP-version ; HNCOCA with D decoupling ; 3D sequence with ; inverse correlation for triple resonance using multiple ; inept transfer steps ; ; F1(H) -> F3(N) -> F2(C=O), HMQC(Ca,t1) -> F3(N,t2) -> F1(H,t3) ; ; Uses Fq list for frequency switching on 13C channel ; o2p=176p->55p->176p ; phase sensitive using States-TPPI (t1) ; phase sensitive using States-TPPI (t2) ; using constant time in t2 ; with H-2 decoupling - requires lockswitch ; Ref. S. Grzesiek & A. Bax, J. Magn. Reson. 96, 432 - 440 (1992) ; T. Yamazaki, W. Lee, C.H. Arrowsmith, D.R. Muhandiram & L.E. Kay, J. Am. Chem. Soc. 116, 11655-11666 (1994) ; Based on wa_hncocaDse, modified for cryoprobe by J.D. Gross ; Added 3-9-19 WATERGATE. Replaced simultaneous 13C/15N 180s with ; interleaved element 90Sx-180Ix-90Sx except during CO evolution ; where sim 13C/15N 180 is employed. JDG 7/12/00 ; Uses doubly selective rsnob for COCA transfer--obviates need for BLOCH-SEIGERT ; can be made with Shape Tool, calculate from excitation region ; compensation. REF--JMR B 106 300-303 1995 ; E. Kupce, J. Boyd and I.D. Campbell ; Converted to HN(COCA)CB JDG 4/23/01 ; 111703 D Frueh adding NLS in the Jim Sun fashion, i.e. with vc lists. ;*********NLS*************************************************************************************** ; 111703 D Frueh adding NLS in the Jim Sun fashion, i.e. with vc lists. ; 1) make the sampling schedule with COAST or sampsched2d (rnmrtk): two columns ; 2) convert to vclist use the script sched2vc (vc2sched) to do the conversion ; 3) in xwinnmr put the corresponding vclist ;IMPORTANT: td1*td2=4*(number of hypercomplex points sampled) ; : first entry in vclist is for 15N ;if 2d.sched has N pairs of points, the vclist will have 2N entries (one for each dimension) ;and the total number of FIDs recorded will be 4N (=td1*td2). ;DATA is stored in the order: ; 1) Re(15N) Re(13C) 2) Im(15N) Re(13C) 3) Re(15N) Im(13C) 4) Im(15N) Im(13C) ; The value of the evolution time of 15N is read first (first entry in vclist) ; Then the value of the 13C evolution (2nd entry in vclist) ;However it is the list that dictates which dimension is labeled first (i.e. which evolution ;periode is varied first). This can be modified if desired from 2d.sched by doing: ;"sort -n -k2 -k1 2d.sched > out.sched" or "sort -n -k1 -k2 2d.sched > out.sched" ;or simply make another list with rnmtk's sampsched2d ; check however that the first entry (and odd entries) correspond to nitrogen: ; example a vc list that looks like: ; 0 -> 15N ; 0 -> 13C (first 4 FIDs :ReRe ReIm ImRe ImIm have been acquired) ; 0 -> 15N ; 1 -> 13C (first 8 FIDs :ReRe ReIm ImRe ImIm have been acquired) ; 0 ; 3 ; 0 ; 5 ;means that 13C is the "fast dimension" (the one incremented first) (see, the value associated ;with 13C changes, whereas those with 15N will only change later on) ; ; a vc list that looks like: ; 0 ; 0 ; 1 ; 0 ; 2 ; 0 ; 7 ; 0 ;would mean that one increments first the 15N dimension ;also check that the sizes are corresponding to what you want (ex. 32 Complex points in 15N,64 Complex points in 13C : ; the list should have values at 63, or 62 for even lines, and not go higher than 32 for odd lines ;(not counting the comments!!!) ;****end****NLS************************************************************************************* ;note: interleaved 90Sx,180Ix,90Sx not necessary for recent cryo-spectrometers or spectrometers without cryoprobes ; or spectrometers without cryoprobes. Then, replace by (CEN_CN2 p2*2 ph10):f2 (p3*2 ph10):f3 ;or on 3.1 and higher by (center (p2*2 ph10):f2 (p3*2 ph10):f3) etc. ;also you might want to replace the gron/groff commands by p16:gp1 etc. ;the current version works on both the oldest and the most recent instruments in our lab. ; on latest Bruker equipement, the LOCKDEC_ON & LOCKDEC_OFF cmds in the beginning and the end ;should be replaced by those currently commented in the pp. #include #include #include #define SED_ON (5u pl15 2u cpds8):f2 #define SED_OFF (2u do 5u pl7):f2 #define WALTZ_ON (5u pl12 p12 ph6 3u 20u cpds1 ph10):f1 #define WALTZ_OFF (2u do p12 ph7 5u pl1):f1 "d0=3u" "d2=2.3m" "d3=5.5m" "d4=12.4m" "d7=4.1m" "d9=12.0m" "d10=3u" "d5=d3-3u-3u-p11" "d20=d4" "d14=d3-5u-p12-3u" "d16=(d19-p3*2)/2" "d17=d4-d3-2u-p12" "d24=d2-50u-d25-200u-d19*1.685" "DELTA=d2-5u-3u-d27-200u" "DELTA2=d9-d3-20u+p7" "DELTA4=d7-p5/2-d27-200u-3u-3u" "DELTA5=6.8m" "CEN_HN1=(p3-p1)/2" "CEN_HN2=(p3-p1)" "l0=(td1*td2/4)" "l10=(td2/2)" aqseq 321 1 30m ze 30m LOCKDEC_ON ; enables the use of the 2H-switch 30m pl31:f3 pl17:f4 2 30m do:f3 H2_LOCK ; 2H-switch to lock, 15N decoupling off 6m LOCKH_OFF ; lockhold off ==> lock active 3 20u*2 10u 4 20u*2 5 20u 6 d1 pl1:f1 1m fq1:f2 ;Set carrier to 176ppm 20u UNBLKGRAD 50u LOCKH_ON ; lockhold on ==> lock not active 20u H2_PULSE ; 2H-switch to pulse 5u pl2:f2 ; Destroy magnetization derived 5u pl3:f3 ; from 13CO and 15N Zeeman order (p2 ph10):f2 3u (p3 ph10):f3 3u gron0 d25 groff 400u (p1 ph10):f1 5u pl3:f3 3u gron27 ; gradient on d27 ; sandwich gradient duration 200u groff ; gradient off DELTA (CEN_HN2 p1*2 ph10):f1 (p3*2 ph10):f3 5u 3u gron27 ; gradient on d27 ; sandwich gradient duration 200u groff ; gradient off DELTA (p1 ph6):f1 ;----------------------------------HzNz---------------------------- 4u pl11:f1 ; power for H2O pulse (p11 ph25:r):f1 ; selective H2O pulse 3u 2u gron29 ; gradient on d27 ; crushing gradient duration 200u groff ; gradient off ;---------------------------------Nz->CO TFER---------------------- (p3 ph8):f3 d14 ; 1/2JHN to start the 1H dec. WALTZ_ON DELTA2 pl7:f2 (p7*0.5 ph10):f2 ; Interleaved 180 (p3*2 ph10):f3 (p7*0.5 ph10):f2 d9 (p3 ph10):f3 ;--------------------------------NzCOz------------------------------- WALTZ_OFF 3u pl2:f2 2u gron28 ; gradient on d27 ; crushing gradient duration groff ; gradient off 200u ;--------------------------------COCA TFER---------------------------- (p2 ph1):f2 3u pl5:f2 3u gron19 ; gradient on d27 ; sandwich gradient duration 200u groff ; gradient off DELTA4 (p5:sp5 ph10):f2 ; doubly selective rsnob 3u pl2:f2 3u gron19 ; gradient on d27 ; sandwich gradient duration 200u groff ; gradient off DELTA4 (p2 ph6):f2 ;---------------------------------COzCAzNz-------------------------------- 2u gron21 ; gradient on d27 ; crushing gradient duration groff ; gradient off 200u fq1:f2 ; Switch 13C carrier to 43ppm ; 1m LOCKDEC_ON WALTZ_ON 2u cpd4:f4 ; 2H decoupling on 4u pl8:f2 ; Null at 176 ppm ;--------------------------------CACB TFER--------------------- (p8 ph9):f2 ; alternate +-x ; Null at +-133ppm DELTA5 pl9:f2 (p9 ph10):f2 ; Null at +-133ppm DELTA5 pl8:f2 ;-------------------------------END CACB TFER------------------------ (p8 ph11):f2 ; y d0 ;CB evolution d0 (p8 ph21):f2 ;END CB evolution ;-------------------------------CACB TFER-------------------------------- DELTA5 pl9:f2 (p9 ph20):f2 DELTA5 pl8:f2 (p8 ph20):f2 ;-------------------------------END CACB TFER--------------------------- ;---------------------------------COzCAzNz------------------------------- WALTZ_OFF 3u do:f4 ; 2H decoupling off ;1m LOCKDEC_OFF 2u gron23 ; gradient on d27 ; crushing gradient duration groff ; gradient off 200u fq1:f2 ; Switch 13C carrier back to 176 ppm ;-----------------------------------COCA TFER------------------------- (p2 ph6):f2 3u pl5:f2 3u gron31 ; gradient on d27 ; sandwich gradient duration 200u groff ; gradient off DELTA4 (p5:sp5 ph10):f2 ; Doubly selective rsnob 3u pl2:f2 3u gron31 ; gradient on d27 ; sandwich gradient duration 200u groff ; gradient off DELTA4 (p2 ph10):f2 ;-----------------------------------COzNz----------------------------- 3u 2u gron18 ; gradient on d30 ; crushing gradient duration groff ; gradient off 200u WALTZ_ON ;-----------------------------------N CT evolution CO-N TFER--------------- (p3 ph3):f3 SED_ON d20 SED_OFF (p7*0.5 ph10):f2 (p3*2 ph4):f3 (p7*0.5 ph10):f2 SED_ON (d17 d10 2u do p12 ph7 d5 3u pl11 p11 ph26 3u pl1):f1 (d4 d10):f2 ; WALTZ_OFF included SED_OFF ;----------------------------------End 15N CT period (CEN_HN1 p1 ph10):f1 (p3 ph10):f3 ;----------------------------------3-9-19 WATERGATE--------------- 4u d24 50u gron25 ; gradient on d25 ; watergate gradient duration groff ; gradient off 200u p1*0.2308 ph6 d19 p1*0.6923 ph6 d19 p1*1.4615 ph6 d16 (p3*2 ph10 ):f3 d16 p1*1.4615 ph7 d19 p1*0.6923 ph7 d19 p1*0.231 ph7 50u gron25 ; gradient on d25 ; watergate gradient duration groff 200u d24 pl31:f3 ; Set power for 15N decoupling 4u BLKGRAD ; Sampling period if (l20 == 1) { go=2 ph31 cpd3:f3 d11 do:f3 wr #0 } if (l20 == 3) { go=2 ph31 cpd3:f3 d11 do:f3 wr #0 if #0 ip3 zd H2_LOCK;15N 20u LOCKH_OFF 20u lo to 3 times 2 20u ip20 20u ip21 lo to 5 times 2 ;13C ; reseting everything 20u rd10 20u rd20 20u rd0 20u rp31 20u rp21 20u rp20 20u rp3 ; increment 15N States-TPPI 95 10u 20u id10 ip3*2 20u dd20 ip31*2 lo to 95 times c 20u ivc ; increment 13C states-TPPI 99 20u id0 ip31*2 20u ip20*2 20u ip21*2 lo to 99 times c 20u ivc lo to 6 times l0 } ;Original linear version ; go=2 ph31 cpd3:f3 ; 30m do:f3 wr #0 if #0 ip3 zd H2_LOCK ; 3m LOCKH_OFF ; 3m ; lo to 3 times 2 ; 10u ; 20u id10 ; 20u dd20 ip31*2 ; lo to 4 times l10 ; 20u rd10 ip20 ; 20u rd20 ip21 ; lo to 5 times 2 ; 20u id0 ip31*2 ; lo to 6 times l0 30m LOCKDEC_OFF ; disables the use of the 2H-switch 20u do:f1 20u do:f2 20u do:f3 exit ph1=0 2 ph9=0 0 2 2 ph3=0 ph4=0 0 0 0 2 2 2 2 ph5=0 ph6=1 ph7=3 ph8=0 0 0 0 2 2 2 2 ph10=0 ph11=1 ph20=0 ph21=1 ph25=0 ph26=2 ph31=0 2 2 0 2 0 0 2 ;-----------PARAMETERS--------- ;------Pulses and powers------- ;------Pulses and powers------- ;p1: 90-deg 1H pulses @ pl1 ;p2: 90-deg 13C-pulses @ pl2 [63u] ;p3: 90-deg 15N-pulses @ pl3 ;p5: 180-deg pulse COCA rsnob ;p7: 180-deg 13C-pls @pl7 [57u] ;p8: 90-deg 13C-pls @pl8 n133ppm [58u] ;p9: 180-deg 13C-pls @pl9 n133ppm [51.8u] ;p11: water selective pulse @ pl11 ~2ms ;pl1: 1H high power level ;pl2: 13Ca 90 pwr lvl (null 121p) ;pl3: 15N high power level ;pl5: pwr for COCA rsnob ;pl6: pl6=pl7 ;pl7: 13C 180 pwr lvl (null 121p) ;pl8: 13C 90 pwr lvl (null 133p) ;pl9: 13C 180 pwr lvl (null 133p) ;pl11: water selective power ;sp5: sp5=pl5 ;spnam5: doubly COCA sel rsnob ;spnam6: 180-deg CA pulse off-res., shf +121p ;------decoupling------------- ;pcpd1: 90-deg pulse for 1H cpds1-dec. @ pl12 ;p12: p12=pcpd1 ;pcpd2: 180-deg pulse for Ca cpds8-dec. @ pl15 ;pcpd3: 90-deg pulse for 15N cpd3-dec. @ pl31 ;pl12: power level for cpds1-dec. on f1 ;pl31: power level for cpd3-dec. on f3 ;pl17: power level for cpd4-dec. on f4 ;cpdprg1: decoupling sequence on f1 [waltz16] ;cpdprg3: decoupling sequence on f3 [garp] ;cpdprg4: decoupling sequence on f4 [waltz16] ;pl15: power level for seduce dec. ;sp15: sp15=pl15 ;spnam15: seduce shape pls for CA dec. shft -121p ;cpdprg8: seduce dec. seq. on f2 for [seduce_wa] ;------delays------------------ ;d0: 13C evolution time ;d2: 2.3 m < 1/4JHN ;d3: 1/2JHN to start the dec. [5.5msec] ;d4: 15N constant time [12.4msec] ;d10: 15N evolution time ;d19: WTG delay [210us] ;------gradients--------------- ;d25: 1m WTG gradient ;d27: 0.5m gradient duration ;d30: 0.3m gradient duration ;gpz18: 30% ;gpz19: 20% ;gpz21: -42% ;gpz22: 12% ;gpz23: 24% ;gpz24: 50% ;gpz25: 50% WTG gradient ;gpz27: 6% ;gpz28: 4% ;gpz29: 60% ;gpz30: 3% ;gpz31: 1.2% ;------others----------------- ;in0: inc. 13C dim. in0: 1/(2*SW(X)) = DW(X) ;in10: inc. 15N dim. in10: 1/(2*SW(X)) = DW(X) ;in20: in10=in20 ;l0: # of points = td1*td2/4 ;l20: 1 for 1D, 3 for 3D ;nd0: 2 ;nd10: 2 ;ds: 32 ;ns: 8 ;phcor25:0