UNDER CONSTRUCTION!



Triple resonance pulse programmes for13C,15N labelled proteins


The pulse programs (Bruker DRX format) presented here are triple resonance experiments for the assignment of 13C,15N labeled proteins. Most of them incorporate a heteronuclear gradient echo for coherence selection for the 15N --> 1H transfer and are combined with sensitivity enhancement and water- flip-back where appropriate.

See the following reference for a review:

M. Sattler, J. Schleucher and C. Griesinger. (1999) Prog. NMR Spectrosc.34, 93-158.


Pulse Programmes

Experiment
 Name
 Info
HNCACB

CBCA(CO)NH

CC(CO)NH-TOCSY

H(CCCO)NH-TOCSY

HCCH-TOCSY

1H-15N HSQC

1H-13C HSQC








Convetions used in the pulse programmes

Rectangular Pulses

Frequency  Flipangle  Name  Power Level  Decoupler
Pulse 		 Decoupling
Power Level 		 TOCSY
Power Level 
1H  90/180  p1/p2  pl1:f1  pcpd1  pl11:f1 (cpd1:f1)  pl21:f1 
13C  90/180  p3/p4  pl2:f2  pcpd2  pl12:f2 (cpd2:f2)  pl22:f2 
15N  90/180  p5/p6  pl3:f3  pcpd3  pl13:f3 (cpd3:f3)  pl23:f3 
2H  - -  -  pcpd4  pl14:f4 (cpd4:f4)  - 

Shaped Pulses

Name  Power Level  Offset  Shape  Flipangle  Duration [us] (500/600MHz) 
p13  sp1  on resonance  G4  90 deg  400 / 400 
p13  sp2  on resonance  G4tr  90 deg  400 / 400 
p14  sp3  on resonance  G3  180 deg  268 / 250 
p14  sp4  +/- offset  G3  180 deg  268 / 250 
p13  sp5  +/- offset  G4  90 deg  400 / 400 
p13  sp6  +/- offset  G4tr  90 deg  400 / 400 
p24  sp8  on resonance  Reburp  180 deg  2ms (CA selective) 
p27  sp11  on resonance  Gauss  270 deg 2.5ms (H2O selective) 

Offset is the difference between the centers of the CO chemical shifts (175ppm) and the CA (55ppm) or aliphatic carbon (40ppm) chemical shifts (120/135ppm).
Pulses that are applied to compensate for Bloch-Siegert phase shifts are denoted BSP.