The multidimensional NMR experiments specifically designed for ²H,¹³C,¹⁵N-labeled proteins are simply modifications of the already existing methodologies, in which the following features are included:

• Increased recycle time (about 2 seconds).
• High levels of ²H in the CA and CB sites allow the use of constant-time (CT) carbon evolution periods with deuterium decoupling resulting in much better resolution in the carbon dimension.

A list of published NMR methods:

• 3D backbone

 

NMR experiment	References	Level of deuteration	Observations
3D HNCA	94JACS6464 94JACS11655	Perdeuterated or 70% 2H-labeled
3D HN(CO)CA	94JACS11655 96JACS6570	perdeuterated or 70% 2H-labeled
3D HN(CA)CB	94JACS11655 96JACS6570	Perdeuterated or 70% 2H-labeled
3D HN(COCA)CB	94JACS11655 96JACS6570	Perdeuterated or 70% 2H-labeled
3D HN(CA)CO	96JMRB112-110 96JMRB194-111	Perdeuterated
3D HBHA(CO)NH	99JMR285
3D HN(COCA)NH	95JB323-5 96JMRB194-111	Perdeuterated
3D HACAN, HACACB,  HACACO, HACA(N)CO	97JACS872	Selective 1HA-labeled  deuterated proteins

 

 
• 3D backbone side-chain:

 

NMR experiment	References	Level of deuteration	Observations
3D C(CC)(CO)NH	95JACS4187	Perdeuterated or 70% 2H-labeled

 

• 4D experiments

 

NMR experiment	References	Level of deuteration	Observations
4D HNCACB	94JACS11655	Perdeuterated or 70% 2H-labeled
4D HN(CA)NH	97JMR214-124
4D HN(COCA)NH	93JACS4369 95JB323-5	Random fractional  deuteration
4D [HBCB/HACA]NNH	96JACS407	Random fractional  50% deuteration
4D [HBCB/HACA](CO)NNH	96JACS407	Random fractional  50% deuteration
4D HCC(CO)NNH	96JACS407	Random fractional  50% deuteration

 

 
• NOE experiments:

 

NMR experiment	References	Level of deuteration	Observations
3D 15N-separated	96JACS407	Perdeuterated or 70% 2H-labeled
3D 13C-separated NOESY-HSQC	96JACS407
4D NN-NOESY  (HSQC-NOESY-HSQC)	95JACS9592 95JACS9594

 

• Experiments for selective amino-acid type assignment and identification :

 

NMR experiment	References	Level of deuteration	Observations
2D beta-carbon-edited HN(COCACB)	96JMRB95-112
3D HN(COCACB)CG	98JMR350-134
3D HN(CACB)CG	98JMR350-134
2D NH2-filetred HSQC 3D H2N(CO)Cb/g 3D H2N(COCb/g)Cb/a	96JB59		Glu and Asn side-chain  amide NH/15N groups
2D refocused H(Ne/f)Cd/e 2D H(Ne/fCd/e)Cg/d	96JB59		Arg side-chain  amide  NH/15N groups and non refocused versions for Lys side-chain amide groups
3D (HM)CMC(CM)HM	98JACS11738 02JB289 03JACS13868	1H-methyl   deuterated proteins	correlating methyl chemical shifts  with CB (val), CG (Leu and Ile)
3D (H)C(CA)NH-COSY	03JACS5701	1H-methyl   deuterated proteins	correlating methyl chemical shifts  with Ile d1 methyl groups
Methyl TROSY	03JACS10420 03MRC843	1H-methyl   deuterated proteins

 
97JMB1223-267
96JMB627-263
95JB339
2D 15N,NH correlations of residues that follow methyl-containing amino acids ( 97JB283 ). specific aa experiment