NMR Experiments and Sequences
Triple Resonance Experiments for Backbone Assignments

For large proteins (> 20 KD), two options are available; TROSY detection and 2H decoupling (requiring 2H labeled proteins). Bruker provides different pulse sequences for these options with their own name conventions; tr (for TROSY) or 2h (for 2H decoupling).
When phase sensitive mode using Echo/Antiecho-TPPI gradient selection (t2) is applied, aqseq 321 is used.

NMR Experiments Bruker Sequences
HNCO
hncogp3d
Acquires 3D; F3(HN) - F2(N) - F1(CO)
trhncoetgp3d
HNCA
hncagp3d
Acquires 3D; F3(HN) - F2(N) - F1(CA)
hncagp2h3d
trhncagp3d2 and trhncagp3d2.2
trhncagp2h3d2; uses INEPT
Caution: HMQC version
trhncagp3d and trhncagp3d.2
trhncagp2d3d
HN(CO)CA
hncocagp3d
Acquires 3D; F3(HN) - F2(N) - F1(CA)
cnst23 = o2p for F1(CA) acquisition
hncocagp2h3d
trhncocaetgp3d
trhncocaetgp2h3d
HN(CA)CO
hncacogp3d
Acquires 3D; F3(HN) - F2(N) - F1(CO)
hncacogp2h3d
trhncacoetgp3d
trhncacoetgp2h3d
CBCA(CO)NH
cbcaconhgp3d
Acquires 3D; F3(HN) - F2(N) - F1(CBCA)
cbcaconhgp2h3d
trcbcaconhetgp3d
trcbcaconhetgp2h3d
HN(CO)CACB
hncocacbgp3d
Acquires 3D; F3(HN) - F2(N) - F1(CBCA)
This is the same as cbcaconhgp3d and it is used for deuterated proteins.
hncocacbgp2h3d
trhncocacbetgp3d
trhncocacbetgp2h3d
HNCACB
hncacbgp3d
Acquires 3D; F3(HN) - F2(N) - F1(CBCA)
hncacbgp2h3d
trhncacbetgp3d
trhncacbetgp2h3d
HBHA(CBCACO)NH
hbhaconhgp3d
Acquires 3D; F3(HN) - F2(N) - F1(HBHA)
HBHA(CBCA)NH hbhanhgp3d
Acquires 3D; F3(HN) - F2(N) - F1(HBHA)
HN(CA)NNH
Connecting Ni-1, Ni, Ni+1
hncannhgp3d, hncannhgpwg3d
Acquires 3D; F3(HN) - F2(N) - F1(N)
hncannhgp2h3d, hncannhgpwg2h3d
trhncannhgp3d, trhncannhetgp3d
trhncannhgp2h3d, trhncannhetgp2h3d
The default setup (d27=12.5ms) shows only Ni-1 and Ni+1
To see Ni, use d27=6.25ms; in Topspin2, d22 acts like d27.
H(NCA)NNH
Connecting HNi-1, HNi, HNi+1
hncannhgp3d.2, hncannhgpwg3d.2
Acquires 3D; F3(HN) - F2(N) - F1(HN)
hncannhgp2h3d.2, hncannhgpwg2h3d.2
trhncannhgp3d.2, trhncannhetgp3d.2
trhncannhgp2h3d.2, trhncannhetgp2h3d.2
The default setup (d27=12.5ms) shows only Ni-1 and Ni+1
To see Ni, use d27=6.25ms; in Topspin2, d22 acts like d27.
HN(CA)HA hncah3d
Acquires 3D; F3(HN) - F2(N) - F1(HA)
HCA(CO)NH hcaconhgp3d
Acquires 3D; F3(HN) - F2(N) - F1(HA)
HCACO hcacogp3d
Acquires 3D; F3(HA) - F2(CA) - F1(CO)

Protein Backbone and Coupling Constants


Protein 13C NMR Spectrum


Triple Resonance Experiments for Backbone Assignments