NMR Experiments for Proteins

NMR Experiments for Proteins

Many versions of multidimensional NMR experiments have been proposed. However. most of them rely on a few general ideas:

2D, 3D and 4D versions of the same experiment, as a function of the variable evolution periods

Most of mutidimensional experiments are based on the HSQC pulse scheme. Thus, conventional modifications of the basic 2D HSQC pulse train can be extrapolated to other 3D and 4D analogues. For instance, the incorporation of the PEP methodology, the use of pulsed field gradients for coherence selection (echo-antiecho) and for solvent suppression (for instance, the water flip-back or the WATERGATE block).

Similar experiments using out-and-back or straight-trough pathways, as a function of the starting and detected spin nuclei.

Substitution of the reverse INEPT by a TROSY block

Incorporation of 2H-decoupling when working on 2H-labeled proteins

Use of constant-time periods and concatenated building blocks, basically by two different finalities: evolution/decoupling of specific J coupling constants and minimization of relaxation losses.

Minimal modifications to obtain specific NMR experiments to measure scalar or dipolar coupling constants, dynamic aspects such as chemical exchange, cross-correlation effects or relaxation rates.

4D	3D	2D	Related experiments	Other versions	J	NOE
HNCACO	HN(CA)CO H(N)CACO	HN(CACO) H(NCA)CO	4D HNCO(-i)CA(i) 3D (HCA)CONH		HN(CA)CO-E.COSY H(N)CA,CO-E.COSY
HNCOCA	HN(CO)CA H(N)COCA HN[COCA]	HN(COCA) H(NCO)CA	4D HNCO(-i)CA(i) 3D (HACA)CO,NH 3D (HA)CA(CO)NH	3D H₂N(CO)CA (Asn and Gln)
HNCAHA	HN(CA)HA H(N)CAHA (H)NCAHA	HN(CAHA) H(NCA)HA		(H)NCAHA-E.COSY 2D arg-H(NC)H
HACANH	HA(CA)NH (HA)CANH HACA(N)H HA[CAN]NH	HA(CAN)H (HACA)NH		HNCAHA	(H)CANNH-E.COSY
HN(CO)CAHA	HN(COCA)HA		(H)N(COCAHA)-TOCSY
HACA(CO)NH	(HACA)CONH (HA)CA(CO)NH HA(CACO)NH [HACA](CO)NH	(HACACO)NH	(H)CA(CO-TOCSY)NH (H)C(C)NH (HACA)CO,NH
(H)CACOCANH	(HCA)CO(CA)NH [HCA][CO(CA)N]H HA(CACO)NH [HACA](CO)NH	(HACACO)NH	(H)CA(CO-TOCSY)NH (H)C(C)NH (HACA)CO,NH
HN(COCA)NH	(H)N(COCA)NH (HNCO)CANH [HN](COCA)NH		(H)N(CO-TOCSY)NH
HN(CA)NH	HN(CA)NNH H(NCA)NNH	HN(CAN)NH (HNCA)NH		3D HN(CA)N HNN HN(C)N
HCACON	HCA(CO)N H(CA)CON	HA(CACO)N HACA(CON)	3D (CO)N(CO)CAH 3D HACAN
HNCACB	HN[CACB] HN(CA)CB	HN(CACB) H(N)CACB	HNCACBCG CB(CA)NH		3D HN(CA)CB-E.COSY
HCCH-TOCSY	H(C)CH-TOCSY (H)CCH-TOCSY	[HC](C)H-TOCSY	3D (H)CCH₃-TOCSY 3D H(C)CH₃-TOCSY	2D HCCH-TOCSY	3D HCC-TOCSY-CCH E.COSY
HCCH-COSY	H(C)CH-COSY			2D and 3D HCCH-E.COSY 2D and 3D CCH-E.COSY
HC(CCO)NH	(H)CC(CO)NH H(CC)(CO)NH (H)CCCO(N)H	(HCCCO)NH (H)CC(CON)H H(CCCON)H	CC(CO)NH (H)C(C)NH H(CC)NH HBHA(CO)NH HBHANH
HCCNNH (HCNH-TOCSY)			4D [HAHB][CACB]NH)
	HNCO	H(N)CO HN(CO) H[NCO]	3D HN(CO)CA H(N)CZ H₂NCO (asn and gln)		HNCO-E.COSY
	HNCA	H(N)CA HN(CA) H[NCA]	3D HN(CO)CA 3D (HA)CANH		HNCA-E.COSY
	HCACO	H(CA)CO HCA(CO)	HCACO-COSY HCACO-TOCSY COCAH HCACO-CBHB	HACACB
	HCAN	HA(CA)N
	HN(CA)N
	(H)NNH-TOCSY		HNN
	CBCA(CO)NH CBCACO(N)H	(CBCACO)NH CBCA(CON)H	(H)CBCA(CO-TOCSY)NH (H)C(CCO)NH HBHA(CO)NH [HBHA-CBCA](CO)NH
	CBCANH	(CBCA)NH CBCA(N)H	[CBCA]NH CCANH CCNH
	HN(CO)CACB HN(COCA)CB	HN(COCACB) H(NCO)CACB H(NCOCA)CB	H₂N(CO)CACB (asn and gln)	HN(CO)CACBCG
	HBHA(CO)NH	(HBHACO)NH HBHA(CON)H	3D H(CCCO)NH [HBHA-CBCA](CO)NH HBHANH
		HSQC	HMQC TROSY	NH2-HSQC Edited-HSQC Relaxation: T1(HN) T2(HN) NOE(HN) T1ro(HN)	HSQC-IPAP
¹³C-¹³C HMQC-NOESY-HMQC HSQC-NOESY-HSQC HSQC-NOESY-TROSY TROSY-NOESY-TROSY	w1-¹³C-edited NOESY-¹³C-HSQC w2-¹³C-edited ¹³C-HSQC-NOESY	w1,w2-¹³C-edited NOESY
¹³C-¹⁵N HMQC-NOESY-HMQC HSQC-NOESY-HSQC HSQC-NOESY-TROSY TROSY-NOESY-TROSY	w1-¹³C-edited NOESY-¹⁵N-HSQC w2-¹⁵N-edited ¹³C-HSQC-NOESY	w1-¹³C-w2-¹⁵N-edited NOESY
¹⁵N-¹³C HMQC-NOESY-HMQC HSQC-NOESY-HSQC HSQC-NOESY-TROSY TROSY-NOESY-TROSY	w1-¹⁵N-edited NOESY-¹³C-HSQC w2-¹³C-edited HSQC-¹⁵N-NOESY	w1-¹⁵N-w2-¹³C-edited NOESY
¹⁵N-¹⁵N HMQC-NOESY-HMQC HSQC-NOESY-HSQC HSQC-NOESY-TROSY TROSY-NOESY-TROSY	w1-¹⁵N-edited NOESY-¹⁵N-HSQC w2-¹⁵N-edited HSQC-¹⁵N-NOESY	w1,w2-¹⁵N-edited NOESY
	NOESY-¹⁵N-HSQC	w2-¹⁵N-edited NOESY
	NOESY-¹³C-HSQC	w2-¹³C-edited NOESY
	¹³C-HSQC-NOESY	w1-¹³C-edited NOESY
	¹⁵N-HSQC-NOESY	w1-¹⁵N-edited NOESY

/td> w2-¹³C-edited NOESY ¹³C-HSQC-NOESY w1-¹³C-edited NOESY ¹⁵N-HSQC-NOESY w1-¹⁵N-edited NOESY