The (+) and (−) enantiomers for a
cryptophane-7-bond-linker-benzenesulfonamide biosensor (C7B) were
synthesized and their chirality confirmed by electronic circular dichroism (ECD)
spectroscopy. Biosensor binding to carbonic anhydrase II (CAII) was
characterized for both enantiomers by hyperpolarized (hp) 129Xe NMR
spectroscopy. Our previous study of the racemic (+/−) C7B
biosensor-CAII complex [Chambers, et al., J. Am. Chem. Soc. 2009,
131, 563–569], identified two “bound”
129Xe@C7B peaks by hp 129Xe NMR (at 71
and 67 ppm, relative to “free” biosensor at 64 ppm), which led to
the initial hypothesis that (+) and (−) enantiomers produce
diastereomeric peaks when coordinated to Zn2+ at the chiral CAII
active site. Unexpectedly, the single enantiomers complexed with CAII also
identified two “bound” 129Xe@C7B peaks:
(+) 72, 68 ppm and (−) 68, 67 ppm. These results are consistent with
X-ray crystallographic evidence for benzenesulfonamide inhibitors occupying a
second site near the CAII surface. As illustrated by our studies of this model
protein-ligand interaction, hp 129Xe NMR spectroscopy can be useful
for identifying supramolecular assemblies in solution.