-
Notifications
You must be signed in to change notification settings - Fork 1
/
cest_15N.cw
303 lines (247 loc) · 5.27 KB
/
cest_15N.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
;15N-CEST experiment based on Lakomek sensitivity-enhanced 15N-T1 measurement
;Chris Waudby, June 2016
;
#include <Avance.incl>
#include <Grad.incl>
#include <Delay.incl>
;list of CEST saturation frequencies
;first line in file 'Hz sfo'
;second line in file should be zero, indicating the reference plane
;give values in Hz relative to sfo3
define list<frequency> N15sat = <$FQ3LIST>
;38 points in steps of 45 Hz:
;define list<frequency> N15sat = {0 -810.0 -765.0 -720.0 -675.0 -630.0 -585.0 -540.0 -495.0 -450.0 -405.0 -360.0 -315.0 -270.0 -225.0 -180.0 -135.0 -90.0 -45.0 0.0 45.0 90.0 135.0 180.0 225.0 270.0 315.0 360.0 405.0 450.0 495.0 540.0 585.0 630.0 675.0 720.0 765.0 810.0}
;define list<frequency> N15sat = {0 -1000 0}
;define list<frequency> N15sat = {0}
"in0=inf2*0.5"
"p2=p1*2"
"p22=p21*2"
"d0=10u"
"d11=30m"
"d26=p21-p1"
"DELTA=2.65m"
"DELTA1=2.65m"
"DELTA2=2.65m-200u"
"DELTA3=2.65m-200u"
"DELTA4=p24-p19*0.6366"
"DELTA5=2.65m-p25-200u"
"DELTA7=2.65m"
"l2=1" ; loop counter for saturation list
"l6=td1"
"l3=td2/2"
; used to identify first point as the reference spectrum,
; and to activate temperature compensation for this point
"DELTA6=30u+p2" ; for zero phase correction
;"cnst18=-800"
;"cnst19=8.2"
aqseq 312
"acqt0=0"
1 ze
1m
2 2m do:f3
1m LOCKH_OFF
d11
3 3m
4 1m
5 1m do:f3
10u UNBLKGRAD
;---------temperature compensation and d1 recovery delay---------
; apply temperature compensation for 1st point (reference with no spinlock)
if "l2==1"
{
10u LOCKH_ON
10u fq=cnst19(bf ppm):f1 ; put 1H on amides
10u pl8:f1
d18 cpds1:f1
10u do:f1
10u fq=0:f1 ; put 1H back on water
10u LOCKH_OFF
}
; purge water before recycle delay
10u pl8:f1
2mp ph0
3mp ph1
10u pl1:f1
;10u pl1:f1
;(p1 ph0):f1
;5u
;p16:gp8
;200u
1m BLKGRAD
d1
1m UNBLKGRAD
;------- kill steady state 15N ------------
10u pl3:f3
(p21 ph0):f3
5u
p16:gp9
200u
;------- first INEPT Hz-> 2HxNz -------------------------
(p1 ph0):f1
5u
DELTA gron0 ; soft gradient to prevent radiation damping
5u groff
(center (p2 ph0):f1 (p22 ph0):f3)
5u
DELTA gron0
5u groff
;------- rephase 2HxNz to Nz-----------------------------
(center (p1 ph5):f1 (p21 ph0):f3)
5u
DELTA1 gron1 ; soft gradient to prevent radiation damping
5u groff
(center (p2 ph0):f1 (p22 ph0):f3)
5u
DELTA1 gron1
5u groff
(center (p1 ph2):f1 (p21 ph6):f3) ; phase-cycle Nz, -Nz for Freeman-Hill decay
; purge pulse to kill any residual HzNz
;--------------------------------------------------------
5u
p16:gp6 ; cleaning gradient
200u
;------15N T1 relaxation period--------------------------
5u pl8:f1 pl18:f3
; water purge
2mp ph0
3mp ph1
if "l2==1" goto 77
5u LOCKH_ON
5u N15sat:f3
5u fq=cnst19(bf ppm):f1
d18 cpds1:f1 cw:f3 ph0
5u do:f1 do:f3
5u LOCKH_OFF
5u fq=0:f1
5u fq=0:f3
77 5u
5u
; water purge
2mp ph0
3mp ph1
5u pl1:f1 pl3:f3
5u
p16:gp7 ; cleaning gradient
200u
;goto 999
;---------- Echo-Antiecho encoding------------------------
(p21 ph7):f3
DELTA6 ;compensation for d0 15N evolution
DELTA5
190u
p25:gp5*EA ; 1ms
10u
(center (p2 ph0):f1 (p22 ph9):f3)
10u
p25:gp5*EA*-1 ; 1ms
190u
DELTA5
; t1 evolution ---------------------------------------------
d0*0.5 gron1
5u groff
d0*0.5 gron1*-1
5u groff
(p2 ph0):f1
d0
;--------Rance-Kay transfer back ----------------------------
(center (p1 ph0):f1 (p21 ph8):f3)
DELTA2 gron2
200u groff
(center(p2 ph0):f1 (p22 ph0):f3)
DELTA2 gron2
200u groff
; ---second INEPT ----------------------------------------
(center (p1 ph1):f1 (p21 ph1):f3) ;DOUBLE 90
DELTA3 gron3
200u groff
(center(p2 ph0):f1 (p22 ph0):f3)
DELTA3 gron3
200u groff
d26*0.5 ;d26 = p21 - p1
(p19 ph0:r):f1 ; fine tune p19 for water suppression
245u
DELTA4
(p2 ph0):f1
5u
p24:gp4 ; 201u
200u
999 10u
10u pl31:f3
20u BLKGRAD
go=2 ph31 cpds3:f3
500u do:f3
d11 wr #0 if #0 zd
; loop over CEST frequencies
1m N15sat.inc
1m iu2
lo to 3 times l6
; Echo/Anti-echo
1m ip8*2
1m igrad EA
1m ru2
1m N15sat.res
lo to 4 times 2
; t1 evolution
1m id0
1m ip7*2
1m ip31*2
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
ph9= 1 3
ph31=1 3 3 1
;-------------NOTES----------------------
;o1p = 4.7 ppm
;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)
;d18: relaxation time
;l6: number of CEST frequencies (including reference) (td1)
;l3: number of complex points (td2 / 2)
;cnst19: chemical shift at center of amide region (ppm)
; 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
;pcpd1: 1H 90 for CPD (4.5 kHz at 900 MHz = 55.5 us)
;pl8: 1H decoupling power (4.5 kHz at 900 MHz = 55.5 us)
;15N pulses
;p21 : 90 deg hard 15N pulse @pl3
;pl3 :15N 90 deg
;pl18: 15N spin-lock power
;CPDPRG3: garp (aq 15N decoupling)
;pcpd3: 15N decoupling (200u @pl31)
;pl31: 15N decoupling power
; gradients
;p16: 1000u
;p17: 200u
;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: 77%
;gpz9: 40%
;gpnam4 SINE.10
;gpnam5 SINE.10
;gpnam6 SINE.50
;gpnam7 SINE.10
;gpnam8 SMSQ10.100
;gpnam9 SMSQ10.100