Jb. Tobin et al., LOW-BARRIER HYDROGEN-BONDING IN MOLECULAR-COMPLEXES ANALOGOUS TO HISTIDINE AND ASPARTATE IN THE CATALYTIC TRIAD OF SERINE PROTEASES, Biochemistry, 34(21), 1995, pp. 6919-6924
We present spectroscopic evidence for the presence of low-barrier hydr
ogen bonds (LBHBs) in molecular complexes composed of carboxylic acids
and 1-methylimidazole (1-MeIm) dissolved in aprotic organic solvents.
A plot of the values of the low-field proton NMR chemical shifts vers
us the aqueous pK(a) of the carboxylic acid exhibits a positive slope
for pK(a) values below 2.1 and a negative slope for higher pK(a) value
s. The chemical shifts for protons near the maximum in this plot are 1
8 ppm, similar to that of 18.3 ppm for His(57)-Asp(102) in the protona
ted catalytic triad of chymotrypsin, The chemical shifts for the proto
n bonded to C2 of 1-MeIm in these complexes also vary with the pK(a) o
f the carboxylic acid and reveal a gradual change from neutral, hydrog
en-bonded 1-MeIm in complexes of weaker acids to hydrogen-bonded 1-met
hylimidazolium ion in complexes of stronger acids. The midpoint chemic
al shift for the C2 proton corresponds to a carboxylic aqueous pK(a) o
f about 2.1. FTIR spectra of the 1-MeIm-carboxylic acid complexes in C
HCl3 indicate that hydrogen bonding is strong and that the complexes a
re of three types: (a) neutral complexes with the weaker acids (pK(a)
greater than or equal to 2.2) in which the antisymmetric carbonyl stre
tching frequencies are lowered relative to the free acids and the ethy
l esters of the same acids; (b) ionic complexes of stronger acids (pK(
a) less than or equal to 2.1) in which the carbonyl stretching frequen
cies are slightly lower than those for the tetrabutylammonium salts of
the same acids; (c) ionic complexes of the same acids (pK(a) less tha
n or equal to 2.1) coexisting with type b, in which the carbonyl stret
ching frequencies are intermediate between those for the tetrabutylamm
onium salts (bond order 1.5) and those of the same acids or their este
rs (bond order 2.0). The latter complexes appear to incorporate a low-
barrier hydrogen bond and are presented as models for the protonated t
riad of chymotrypsin and other serine proteases. These enzymes have be
en postulated to utilize a low-barrier hydrogen bond between His(57) a
nd Asp(102) to facilitate the abstraction of the beta-OH proton from S
er(195) in the course of catalysis [Frey, P. A., Whitt, S. A., and Tob
in, J. B. (1994) Science (Washington, D.C.) 264, 1997-1930].