Peptide thioester preparation via intramolecular O-to-S acyl transfer is a recently developed method for protein chemical synthesis through Fmoc chemistry. Theoretical calculations have been carried out to study the mechanism for the formation of thioesters via O-to-S acyl transfer. It is found that the O-to-S acyl transfer occurs via an anionic stepwise mechanism in which the cleavage of the C-O bond is the rate-limiting step. The side reaction of hydrolysis also proceeds through an anionic stepwise process, and its rate-limiting step is the attack of the hydroxide ion on the carbonyl carbon. Increase of the chain length between the ester O atom and the S atom can increase the energy barrier of the O-to-S acyl transfer. On the other hand, substituents at the α-position of the ester can reduce the energy barrier.

A new fluorescent probe for monitoring Zn2+ was synthesized based on the structure of 7-hydroxyquinoline. Compared with 8-substituted quinolines, the new probe exhibited higher selectivity for Zn2+ over Cd2+. Its fluorescence enhancement (14-fold) and nanomolar range sensitivity (Kd = 0.117 nM) were favorable toward biological applications. Experiments also showed that a cell-permeable derivative of the new probe was potentially useful for two-photon imaging in living cells.