;sehCaNH3Dws.jb ; ;hCaNH experiment with ;sensitivity enhanced CaN transfer using TROP pulse and ;sensitivity enhanced NH transfer using TROP pulse ;water suppression after 1st CP ;selective refocusing pulses and Bloch-Siegert shift complensation [Cavanagh: Protein NMR spectroscopy 2nd ed. pp 172] ;JB + ZT Faculty of Science, Charles University in Prague ;MB Technical University of Munchen ; ; ;$CLASS=BioSolids ;$DIM=3D ;$TYPE=Heteronuclear Triple-resonance ;$SUBTYPE=sensitivity enhanced ;$COMMENT=hCaNH sensitivity-enhanced 3D using TROP pulses with water suppression ;Parameters: ;p1 : H 90 at plw1 ;p2 : N 90 at plw2 ;p3 : C 90 at plw3 ;p13 : hCa CP contact time ;p32 : CaN seCP contact time ;p21 : NH seCP contact time ;p33 : CO selective 180 pulse at sp33 (~256 to 275 us at 600 MHz) ; (is linearly proportional to 1/B0! --> adjust length to B0 used)(Q3.2000) ;p34 : Ca selective 180 pulse at sp34 (~256 to 350 us at 600 MHz) ; (is linearly proportional to 1/B0! --> adjust length to B0 used)(Q3.2000) ;pl1 : H hard pulse power ;pl2 : N hard pulse power ;pl3 : C hard pulse power ;pl12 : H high-power decoupling power ;pl16 : N high-power decoupling power ;pl18 : C high-power decoupling power ;pl51 : 1H CW decoupling power during CN seCP (usually not used =0W) ;pl13 : H water-suppression power ;sp3 : H hCa CP (ramp) power ;sp30 : C hCa CP (rect) power ;sp32 : C CaN seCP (TROP) power ;sp37 : N CaN seCP (TROP) power ;sp2 : N NH seCP (TROP) power ;sp20 : H NH seCP (TROP) power ;sp33 : CO selective 180 power ;sp34 : Ca selective 180 power ;d19 : delay for water suppression (~100 to 300 ms) ;d0 : C incremental delay (t1) ;d10 : N incremental delay (t2) ;d1 : recycle delay; 1 to 5 times 1H-T1 ;pcpd1 : H decoupling pulse for cpdprg1 ;pcpd2 : N decoupling pulse for cpdprg2 ;pcpd3 : C decoupling pulse for cpdprg3 ;cpdprg1 : H decoupling file (at pl12) during d0, d10 (e.g. rcw_ApA_pl12) ;cpdprg2 : N decoupling file (at pl16) during acq (e.g. waltz16_pl16) ;cpdprg3 : C decoupling file (at pl18) during acq (e.g. waltz16_pl18) ;cpdprg4 : H water suppression ('cwX_13nofq' at pl13 (15 kHz)) ;cpdprg5 : H water suppression ('cwY_13nofq' at pl13 (15 kHz)) ;spnam3 : ramp90100.1000 (H hCa CP) ;spnam30 : square.100 (C hCa CP) ;spnam32 : TROP-CaN-MASxkHz-x-chan13C.sh (C CaN seCP) ;spnam37 : TROP-CaN-MASxkHz-x-chan15N.sh (N CaN seCP) ;spnam2 : TROP-NH-MASxkHz-x-chan15N.sh (N NH seCP) ;spnam20 : TROP-NH-MASxkHz-x-chan1H.sh (H NH seCP) ;spnam33 : Q3.2000 (CO selective shape) ;spnam34 : Q3.2000 (Ca selective shape) ;spoffs30 : diff. from current carrier to Ca freq. (in Hz, is calculated) ;spoffs32 : diff. from current carrier to Ca freq. (in Hz, is calculated) ;spoffs33 : diff. from current carrier to CO freq. (in Hz, is calculated) ;spoffs34 : diff. from current carrier to Ca freq. (in Hz, is calculated) ;cnst21 : Frequency offset for CO (~174 ppm) ;cnst22 : Frequency offset for Ca (~54 ppm) ;cnst26 : Frequency offset for CO-Ca middle (~114 ppm, is calculated) ;ns : number of scans n x 4 ;td1 : number of C increments ;td2 : number of N increments ;FnMODE : echo-antiecho in both F1 and F2 "cnst26=((cnst21-cnst22)/2)+cnst22" ; center between CO & Ca ; calculation of shape offsets "spoff34=0" "spoff30=spoff34" "spoff32=spoff34" "spoff33=bf3*((cnst21-cnst22)/1000000)" "in0=inf1/2" "in10=inf2/2" "d0=1u" "d10=0u" define delay DELTA "DELTA=d0*2+larger(p1*2,p33)-p33" ; symmetry delay "acqt0=0" ; baseopt correction aqseq 312 1 ze 2 d1 do:f2 do:f3 0.5u fq=cnst21(bf ppm):f3 ; call cnst21 for ased 0.5u fq=cnst22(bf ppm):f3 ; switch to Ca freq. #include ;Max. 50 ms acquisition time ; Initial excitation and hCa CP (p1 pl1 ph0):f1 (p13:sp3 ph1):f1 (p13:sp30 ph2):f3 ; water suppression (p3 pl3 ph20):f3 ; brings magn. to z 0.5u cpds4:f1 d19*0.25 0.5u do:f1 0.5u cpds5:f1 d19*0.25 0.5u do:f1 0.5u cpds4:f1 d19*0.25 0.5u do:f1 0.5u cpds5:f1 d19*0.25 0.5u do:f1 (p3 pl3 ph21):f3 ; brings magn. to xy ; t1 13C evolution with selective refocusing and Bloch-Siegert-shift compensation 1u cpds1:f1 ; 1H decoupling on (p33:sp33 ph14):f3 ;Bloch-Siegert compensation @CO DELTA ; symmetry delay (p34:sp34 ph15):f3 ;Bloch-Siegert compensation @Ca d0 (center (p2*2 pl2 ph11):f2 (p33:sp33 ph16):f3) ; CO selective and 15N refocusing d0 1u do:f1 ; 1H decoupling off ; echo-antiecho determination (p3 pl3 ph3):f3 ; 1st half of 180 (p3 pl3 ph4):f3 ; 2nd half of 180 ; CaN seCP (p32:sp32 ph5):f3 (p32:sp37 ph6):f2 (p32 pl51 ph13):f1 ; t2 15N evolution 0.1u cpds1:f1 fq=cnst26(bf ppm):f3 ; 1H decoupling on 13C freq. to CO-Ca middle d10 (p3*2 pl3 ph12):f3 d10 0.1u do:f1 ; 1H decoupling off ; echo-antiecho determination (p2 pl2 ph7):f2 ; 1st half of 180 (p2 pl2 ph8):f2 ; 2nd half of 180 ; NH seCP (p21:sp2 ph9):f2 (p21:sp20 ph10):f1 ; Acquisition go=2 ph31 cpds2:f2 cpds3:f3 10m do:f2 do:f3 mc #0 to 2 F1EA(calph(ph4, +180),caldel(d0, +in0)) F2EA(calph(ph8, +180) & calph(ph4, +180),caldel(d10, +in10)) HaltAcqu, 1m ;jump address for protection files exit ph0= 1 ; H excitation hard pulse ph1= 0 ; H HC CP ph2= 1 3 ; C HC CP ph20= 0 2 ; water supp. first 90 ph21= 2 0 ; water supp. second 90 ph11= 0 ; N 180 during d0 ph3= 0 ; 1st half of 13C 180 ph4= 2 ; 2nd half of 13C 180 ph5= 0 ; C CN seCP ph6= 0 0 2 2 ; N CN seCP ph13= 0 ; H CW during CN seCP ph12= 0 ; C 180 during d10 ph7= 0 ; 1st half of 15N 180 ph8= 2 ; 2nd half of 15N 180 ph9= 0 ; N NH seCP ph10= 0 ; H NH seCP ph14= 0 ; selective 180 on C ph15= 0 ; selective 180 on C ph16= 0 ; selective 180 on C ph31= 1 3 3 1 ; detection