;sehetnoe_fb.al ;2D 1H/15N heteronuclear NOE with water along +Z ;Farrow, N.A. et al, Biochemistry (1994), 33, 5984-6003. ; ;Written up by Andrew Lee, lee@winnipeg.chem.buffalo.edu ;Implemented and tested on DMX-600 and DMX-750 by B. Volkman, ;volkman@nmrfam.wisc.edu. 7/20/96 ; ; Disclaimer: This pulse program is provided "as is" for your ; information. Support for the use of this pulse program is only ; provided to users of the National Magnetic Resonance Facility ; at Madison (NMRFAM). Users of this pulse program employ it at ; their own risk. Neither NMRFAM or University of Wisconsin-Madison ; are liable for any physical or other damage incurred during the ; use of this pulse program. ;BMRB Pulse Sequence Accession Number: 20 ; ;Suitable grdprog and *.incl files are appended to ;the end of this pulse program, and should be removed and ;placed in appropriate /pp or /gp directory prior to execution ;of this pulse program. ;use gradient program al_hetnoe ; ;**Set these parameters manually** ;RF pulses ;F1= proton at H2O ;F2=15N at amides (~116ppm) ;p1 - 1H high power 90 @ pl1 ;p2 - 15N high power 90 @ pl2 ;p5 - 1H high power 120 @ pl1 ;pcpd2 - 15N aliphatic decoupling 90 @ pl12 ;cpdprg2 - waltz16 or garp ; ;Gradient pulses ;p12=500u ;p17=1.25m ;p18=125u ; ;d0=3u ;IN0=1/SW*2 in 15N dim. ; ;recycle delay = d1+d20+d21 ;l8=600 - for 3s NOE buildup period ;**The following parameters are preset** "d2=2.25m-p12" ;p12=GRADIENT12 pulse length "d3=d2-d16" "d4=2.75m" "d5=500u" "d6=p1" "d7=p2" "d10=5m-p5" "d11=30m" "d12=20u" "d14=d4-d12+p1*2" "d15=d4-d12-p17" ;p17=GRADIENT17 pulse length "d16=300u" "d20=p5*l8" "d21=d10*l8" "d25=d5-p18" "d29=p2-p1" #include #include 1 ze d11 pl1:f1 pl2:f2 ;pl3:f3 2 d11 do:f2 d11 5 d11 d11*2 6 d12 ;****************** reference experiment ********************** d11 d12 LOCKH_OFF d1 pl1:f1 pl2:f2 d20 d21 ;no cpdt in reference expt. d12 ;-------------------------------------------------------- (p2 ph11):f2 d14 ;fq1:f1 d12 (p2*2 ph12):f2 d0 ;t1/2 (p1*2 ph0):f1 d12 LOCKH_ON GRADIENT17(cnst25) ;g1=1.25m @ 30 G/cm for coh. selection d15 d0 ;t1/2 ;---- start RINEPT -------------------------------------- (p2 ph13):f2 d6 (p1 ph0):f1 GRADIENT12(cnst21) ;g2=0.5hm @ 4 G/cm d2 (p2*2 ph0):f2 (p1*2 ph0):f1 d3 GRADIENT12(cnst21) d16 d7 ;---- refocus orthogonal component ---------------------- (p2 ph1):f2 (p1 ph1):f1 GRADIENT12(cnst21) ;g3=g2=0.5hm @ 4 G/cm d2 (d29 p1*2 ph0):f1 (p2*2 ph0):f2 d3 GRADIENT12(cnst21) d16 ;---- put all protons transverse, refocus coherences ---- (p1 ph2):f1 d5 (p1*2 ph0):f1 GRADIENT18(cnst27) ;g4=0.125m @ 27.8 G/cm for refocusing d25 pl12:f2 go=2 ph31 cpd2:f2 d11 do:f2 wr #0 if #0 zd ;******************* NOE experiment ********************* 7 d11 do:f2 d11*4 d11 LOCKH_OFF d1 pl1:f1 pl2:f2 8 (p5 ph0):f1 d10 lo to 8 times l8 d12 ;-------------------------------------------------------- (p2 ph11):f2 d14 ;fq1:f1 d12 pl1:f1 (p2*2 ph12):f2 d0 ;t1/2 (p1*2 ph0):f1 d12 LOCKH_ON GRADIENT17(cnst25) ;g1=1.25m @ 30 G/cm for coh. selection d15 d0 ;t1/2 ;---- start RINEPT -------------------------------------- (p2 ph13):f2 d6 (p1 ph0):f1 GRADIENT12(cnst21) ;g2=0.5hm @ 4 G/cm d2 (p2*2 ph0):f2 (p1*2 ph0):f1 d3 GRADIENT12(cnst21) d16 d7 ;---- refocus orthogonal component ---------------------- (p2 ph1):f2 (p1 ph1):f1 GRADIENT12(cnst21) ;g3=g2=0.5hm @ 4 G/cm d2 (d29 p1*2 ph0):f1 (p2*2 ph0):f2 d3 GRADIENT12(cnst21) d16 ;---- put all protons transverse, refocus coherences ---- (p1 ph2):f1 d5 (p1*2 ph0):f1 GRADIENT18(cnst27) ;g4=0.125m @ 27.8 G/cm for refocusing d25 pl12:f2 go=7 ph31 cpd2:f2 d11 do:f2 wr #0 if #0 zd ;********************************************************* d11 ip13*2 lo to 5 times 2 d11 id0 d11 ip11*2 d11 ip31*2 lo to 6 times l4 d11 LOCKH_OFF exit ph0=0 ph1=1 ph2=2 ph3=3 ph11=1 ph12=0 1 2 3 ph13=0 ph31=0 2 ***** Gradient Program - remove this and following **** lines before executing pulse program ** bvhbhaconhm.r **** loop ns*2 { p17 { (0)+(cnst25*cnst30) | (0) | (0)+(cnst25) } { (0) | (0) | (0) } p12 { (0) | (0) | (0)+(cnst21) } { (0) | (0) | (0) } p12 { (0) | (0) | (0)+(cnst21) } { (0) | (0) | (0) } p12 { (0) | (0) | (0)+(cnst21) } { (0) | (0) | (0) } p12 { (0) | (0) | (0)+(cnst21) } { (0) | (0) | (0) } p18 { (0)+(cnst27*cnst30) | (0) | (0)+(cnst27) } { (0) | (0) | (0) } } loop ns*2 { p17 { (0)+(cnst25*cnst30) | (0) | (0)+(cnst25) } { (0) | (0) | (0) } p12 { (0) | (0) | (0)+(cnst21) } { (0) | (0) | (0) } p12 { (0) | (0) | (0)+(cnst21) } { (0) | (0) | (0) } p12 { (0) | (0) | (0)+(cnst21) } { (0) | (0) | (0) } p12 { (0) | (0) | (0)+(cnst21) } { (0) | (0) | (0) } p18 { (0)+(cnst28*cnst30) | (0) | (0)+(cnst28) } { (0) | (0) | (0) } } ;Gradal.incl - include file for Gradient Spectroscopy ;avance-version ;version 94/06/27 #define GRADIENT10(ampl) p10:ngrad:c34 #define GRADIENT11(ampl) p11:ngrad:c34 #define GRADIENT12(ampl) p12:ngrad:c34 #define GRADIENT13(ampl) p13:ngrad:c34 #define GRADIENT14(ampl) p14:ngrad:c34 #define GRADIENT15(ampl) p15:ngrad:c34 #define GRADIENT16(ampl) p16:ngrad:c34 #define GRADIENT17(ampl) p17:ngrad:c34 #define GRADIENT18(ampl) p18:ngrad:c34 #define GRADIENT19(ampl) p19:ngrad:c34 #define GRADIENT20(ampl) p20:ngrad:c34 #define BLKGRAD setnmr2^0 setnmr0^34 #define UNBLKGRAD setnmr2|0 setnmr0|34 #define BLKGRAMP setnmr0^34 #define UNBLKGRAMP setnmr0|34