An Antibody-Catalyzed Isomerization Reaction

Monoclonal antibodies (mAbs) were generated against the coplanar transition state (TS ⧧ ) analogue 1 and assayed for their ability to catalyze the isomerization of bridged biphenyls 4, 6, and 7 . This is a relatively simple unimolecular reaction whose activation barrier arises from unfavorable steric interactions between the two benzylic methylene groups and strain in the bridging ring system. Seven mAbs were found that catalyzed the isomerization of 4 to 6; the most efficient (mAb 64D8E10) has k cat and K M values of 4.3 × 10 -5 s -1 and 420 μM, respectively. This corresponds to a rate enhancement over the unimolecular uncatalyzed reaction ( k cat / k uncat ) of 2900. The dissociation constant for the TS ⧧ analogue, K d, was determined to be 210 nM. For both the antibody (64D8E10) catalyzed and uncatalyzed reactions, the free energy of activation (Δ G ⧧ ) is comprised largely of the enthalpy term; the antibody decreases the enthalpy of activation by 5 kcal/mol. Despite relatively large differences in the values of k cat / k uncat for the five antibodies, the ratios of K d to K M ( 4 ) are very similar. It is likely that the antibodies catalyze this reaction by reducing both ring strain and nonbonded steric interactions in the transition state.