Hydrogen bonds are of fundamental importance in stabilizing biomolecular structures. NMR evidence for the existence of hydrogen bonds in biomolecules ( 99MFC961 ) can be ascertained from:

Water-protein interactions

The effect of deuterium exchange by determining:

Reduced hydrogen exchange rates with the solvent.

Isotope shift effects upon substitution with ²H or ³H.

Amide hydrogen D/H fractionation factor ( 94BIO1029 and 96JACS12864)

Deuterium quadrupolar coupling constants ( 97JMR61-124, 97JMR54-127)

Amide proton temperature coefficients (01JB249-21)

Effect on isotropic ( 83JACS5948, 92JACS4320, 93SCI1491) and anisotropic chemical shifts ( 97JACS8076, 97JACS8985).

Sequential ¹J_CO,N coupling constants ( 95JACS405, 96JACS7859, and 98JB1-11 ).

Scalar coupling constants measured through hydrogen bonds

Cross-correlation between CSA(1H) and DD(NH) ( 01JMR149-149)

On the other hand, the hydrogen bonding pattern can also give useful information about the secondary structure of proteins:

3₁₀-helix: i^__>i+3 hydrogen bonding

Alpha helix: i^__>i+4 hydrogen bonding

Examples:

Temperature-dependence of protein hydrogen bonding parameters as cross-hydrogen scalar couplings 3hJ(NCO), chemical shifts, amide proton exchange rates, 15N relaxation parameters as well as 1J(N-CO) and 1J(NH) coupling constants (02JMB739).

Hydrogen bonds in serine protease active sites (01MRCS199)