-
Notifications
You must be signed in to change notification settings - Fork 1
/
Copy pathT1rhoN15_hsqc.cw
313 lines (264 loc) · 6.12 KB
/
T1rhoN15_hsqc.cw
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
128
129
130
131
132
133
134
135
136
137
138
139
140
141
142
143
144
145
146
147
148
149
150
151
152
153
154
155
156
157
158
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
174
175
176
177
178
179
180
181
182
183
184
185
186
187
188
189
190
191
192
193
194
195
196
197
198
199
200
201
202
203
204
205
206
207
208
209
210
211
212
213
214
215
216
217
218
219
220
221
222
223
224
225
226
227
228
229
230
231
232
233
234
235
236
237
238
239
240
241
242
243
244
245
246
247
248
249
250
251
252
253
254
255
256
257
258
259
260
261
262
263
264
265
266
267
268
269
270
271
272
273
274
275
276
277
278
279
280
281
282
283
284
285
286
287
288
289
290
291
292
293
294
295
296
297
298
299
300
301
302
303
304
305
306
307
308
309
310
311
; 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"
# else
"DELTA6=30u"
# endif
"cnst21=176"
"cnst22=56"
"cnst18=-800"
"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
;---------temperature compensation---------
# ifdef TEMP_COMPENSATION
"p17=p18+1m-vp"
"d17=d1-p17"
# endif
"d8=p8+vp*0.25-p1*2-2.9u"
"d9=vp*0.5-p1*4-27u"
"d29=p8"
;-------- Temperature compensation and d1 recovery delay----------------
#ifdef TEMP_COMPENSATION
10u fq=cnst18(bf ppm):f3
10u pl8:f3
(p17 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-----------------------------
(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 pl8:f3 ; change to pl0:120dB for Avance I or DRX systems
;------ N15 T1rho relaxation period-----------
# ifdef LABEL_CN
(d8 p1 ph0 3u p1*2 ph1 3u p1 ph0 3u d9*0.5 gron9 3u groff 6u d9*0.5 gron9*-1 3u groff 6u p1 ph0 3u p1*2 ph1 3u p1 ph0):f1 (p8:sp8 ph0 3u pl8 vp ph0 p8:sp9 ph0):f3 (d29 d9 p4*2 ph0 3u 3u pl2 p4*2:sp4 ph0):f2
#else
(d8 p1 ph0 3u p1*2 ph1 3u p1 ph0 3u d9*0.5 gron9 3u groff 6u d9*0.5 gron9*-1 3u groff 6u p1 ph0 3u p1*2 ph1 3u p1 ph0):f1 (p8:sp8 ph0 0.1u pl8 vp ph0 p8:sp9 ph0):f3
#endif
5u
p22:gp8
200u
;---------- Echo- Antiecho encoding------------------------
77 3u pl7:f3
#ifdef LABEL_CN
3u pl4:f2
#else
3u
#endif
3u pl1:f1
(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)
5u
p25:gp5*EA*-1
195u
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 back transfer ----------------------------
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
(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
2m ivp
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
ph2=2
ph5=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
; 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
;p18 maximum duration of spin-lock; temperature compensation
;pl7: 15N 90 deg
;pl8: 15N spin-lock power
;pl0 120 dB
;sp8 @pl8 spin-lock power
;sp9 @pl8 spin-lock power
;spnam8 AHP TanhTan (1st half)
;spnam9 AHP TanhTan (2nd half)
;spoal8 1
;spoal9 0
;CPDPRG3: garp (aq 15N decoupling)
;pcpd3: 15N decoupling (200u @pl31)
;pl31: 15N decoupling power
; gradients
;p20: 1000u
;p21: 200u
;p22: 300u
;p24: 201u Echo/Anti-echo decoding gradient
;p25: 1000u Echo/Anti-echo half-encoding gradient
;for z-only gradients
;gpz0: 3%
;gpz1: 2%
;gpz2: -5%
;gpz3: -0.5%
;gpz4: 40%
;gpz5: 40%
;gpz6: 30%
;gpz7: -50%
;gpz8: 40%
;gpz9: 0.5%
;gpz10:-1.5%
;gpnam4 SINE.10
;gpnam5 SINE.10
;gpnam6 SINE.50
;gpnam7 SINE.10
;gpnam8 SINE.10