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T1N15_hsqc.cw
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T1N15_hsqc.cw
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; 15N-T1 relaxation experiment with sensitivity enhanced (Rance-Kay) HSQC read-out
; relaxation time = 40ms * vclist
; F2 = QF, F1 = Echo-AntiEcho
; for 15N, 2H15N, 15N13C and 2H15N13C labelled proteins
; written by NL 10/27/11
; see footnotes
#include <Avance.incl>
#include <Grad.incl>
#include <Delay.incl>
;#define LABEL_CN ; switch on for 13C labelled samples
#define TEMP_COMPENSATION
"in0=inf1*0.5"
"l3=td1/2"
"l6=td2"
"d0=10u"
"d11=30m"
"d26=p7-p1"
"DELTA=2.65m"
"DELTA1=2.65m"
"DELTA2=2.65m-200u"
"DELTA3=2.65m-200u"
"DELTA4=p24-p19*0.63"
"DELTA5=2.65m-p25-200u"
"DELTA7=2.65m"
"l1=1"
"l2=1"
# ifdef LABEL_CN
"DELTA6=35u+p4*4"
"d25=20m-p15*0.5-p4*4-5u"
# else
"DELTA6=30u"
"d25=20m-p15*0.5"
# endif
"cnst19=8.6"
"cnst21=176"
"cnst22=56"
"cnst18=-800"
"spoff5=(bf1*cnst19)/1000000 - o1"
"spoff4=bf2*((cnst22-cnst21)/1000000)"
"acqt0=0"
1 ze
1m
2 2m do:f3
#ifdef LABEL_CN
10u do:f2
#endif
2m LOCKH_OFF
d11
3 3m
4 1m
5 1m do:f3
1m
#ifdef LABEL_CN
10u do:f2
#endif
1m BLKGRAD
10u pl1:f1
#ifdef LABEL_CN
10u pl4:f2
#endif
;********** echo/anti-echo encoding *******
;;---------temperature compensation and d1 recovery delay---------
# ifdef TEMP_COMPENSATION
"d17=d1-p18"
10u fq=cnst18(bf ppm):f3
10u pl8:f3
(p18 ph0):f3 ; 15N pulse is applied far off-resonance
10u
10u fq=0:f3
d17
# else
d1
# endif
1m UNBLKGRAD
10u pl7:f3
;------- kill steady state 15N ------------
(p7 ph0):f3
5u
p20:gp6
200u
;------- first INEPT Hz-> 2HxNz -------------------------
(p1 ph0):f1
5u
DELTA gron0 ; soft gradient to prevent radiation damping
5u groff
(center(p1*2 ph0):f1 (p7*2 ph0):f3)
5u
DELTA gron0
5u groff
;------- rephase 2HxNz to Nz-----------------------------
(center(p1 ph5):f1 (p7 ph0):f3)
5u
DELTA1 gron1 ; soft gradient to prevent radiation damping
5u groff
(center (p1*2 ph0):f1 (p7*2 ph0):f3)
5u
DELTA1 gron1
5u groff
(p7 ph6):f3 ; phase-cycle Nz, -Nz for Freeman-Hill decay
5u
;--------------------------------------------------------
(p1 ph2):f1 ; purge pulse to kill any residual HzNz
;goto 999
5u
p21:gp7 ; cleaning gradient
100u
100u pl0:f1
;------15N T1 relaxation period--------------------------
if "l2==1" goto 77
70 d25*0.5
# ifdef LABEL_CN
3u pl4:f2
(p4*2 ph0 3u 3u pl2 p4*2:sp4 ph0):f2
# endif
d25*0.5
(p15:sp5 ph0):f1
d25*0.5
# ifdef LABEL_CN
3u pl4:f2
(p4*2 ph0 3u 3u pl2 p4*2:sp4 ph0):f2
# endif
d25*0.5
lo to 70 times c ; delay=c*2*d25 (20ms)
;---------- Echo- Antiecho encoding------------------------
77 10u pl1:f1
#ifdef LABEL_CN
5u pl4:f2
#else
5u
#endif
5u pl7:f3
(p7 ph7):f3
;goto 999
DELTA6;compensation for d0 15N evolution
DELTA5
190u
p25:gp5*EA
10u
(center (p1*2 ph0):f1 (p7*2 ph7):f3)
10u
p25:gp5*EA*-1
190u
DELTA5
; t1 evolution ---------------------------------------------
89 d0*0.5 gron1
5u groff
d0*0.5 gron1*-1
5u groff
# ifdef LABEL_CN
(center (p1*2 ph0):f1 (p4*2 ph0 3u 3u pl2 p4*2:sp4 ph0):f2)
# else
(p1*2 ph0):f1
# endif
d0
;goto 999
;--------Rance-Kay transfer back ----------------------------
98 (center (p1 ph0):f1 (p7 ph8):f3)
DELTA2 gron2
200u groff
(center(p1*2 ph0):f1 (p7*2 ph0):f3)
DELTA2 gron2
200u groff
; ---second INEPT ----------------------------------------
(center (p1 ph1):f1 (p7 ph1):f3) ;DOUBLE 90
DELTA3 gron3
200u groff
(center(p1*2 ph0):f1 (p7*2 ph0):f3)
DELTA3 gron3
200u groff
(d26*0.5 p19 ph0:r):f1
998 245u
DELTA4
(p1*2 ph0):f1
5u
p24:gp4
200u
999 5u ; jump point for water optimisation
#ifdef LABEL_CN
5u pl30:f2 ; 13C DECOUPLING POWER
#else
5u
#endif
10u pl31:f3
20u BLKGRAMP
; acquisition
#ifdef LABEL_CN
go=2 ph31 cpds3:f3 cpd2:f2
500u do:f3
500u do:f2
#else
go=2 ph31 cpds3:f3
1m do:f3
#endif
1m LOCKH_OFF
d11 wr #0 if #0 zd
; loop over relaxation times
1m ivc
1m iu2
lo to 3 times l6
; loop over 15N evolution
1m ip8*2
1m igrad EA
1m ru2
lo to 4 times 2
1m id0
lo to 5 times l3
1m do:f3
1m BLKGRAD
exit
ph0=0
ph1=1 ;check right phase for Boltzmann !!!!!
ph2=2
ph5=1 1 1 1
ph6=1 1 3 3
ph7= 1 3
ph8= 0
ph31=1 3 3 1
;-------------NOTES----------------------
;o1p = 4.7 ppm
;o2p=176 ppm (CO)
;o3p=119 ppm
;NS=4*n
;in0=inf/2
;SW=1/(2*in0)
;echo-antiecho in N15 (process as Complex in NmrDraw before splitting the spectra)
; 1H pulses
;p1: 90 deg hard 1H pulse @pl1
;p19 last 90 deg hard 1H pulse @pl1, can be adjusted for improving water-supression
;pl1: 1H 90 deg
;pl0: 120 dB
;p15: 1618us (@ 700 MHz) 180 deg IBurp2 pulse on 1H (sp5)
;sp5: 180 deg IBurp2 pulse on 1H (p15)
;spnam5: IBurp2
;spoffs5: 8.6 ppm, should be centered in amide region but not touch the water
; 13C pulses
;p4: 13CO selective 180 deg (23.7*2us @ 600 MHz) @pl4
;pl4: 13C 90 deg
;sp4: 13CA selective 180 deg (23.7*2us @ 600 MHz)
;15N pulses
;p7 : 90 deg hard 15N pulse @pl7
;d17: calculated spin-lock delay
;p18: maximum duration of spin-lock; temperature compensation
;pl7 :15N 90 deg
;pl8: 15N spin-lock power
;CPDPRG3: garp (aq 15N decoupling)
;pcpd3: 15N decoupling (200u @pl31)
;pl31: 15N decoupling power
; gradients
;p20: 1000u
;p21: 200u
;p24: 201u Echo/Anti-echo decoding gradient
;p25: 1000u Echo/Anti-echo half-encoding gradient
;for z-only gradients
;gpz10: -1.5%
;gpz2: -5%
;gpz3: -0.5%
;gpz4: 40%
;gpz5: 40%
;gpz0: 3%
;gpz1: 2%
;gpz6: 30%
;gpz7: -50%
;gpnam4 SINE.10
;gpnam5 SINE.10
;gpnam6 SINE.50
;gpnam7 SINE.10