Intermediate Trapping Using Force Spectroscopy

Abstract

Trapping of reaction intermediates has been used as an approach to study the mechanism of various reactions. It serves the purpose of providing a convincing proof as to whether a particular reaction is going via a certain pathway. Here we trap and study an intermediate that is formed during the enzymatic reaction of Sortase A using the help of Force Spectroscopy via Atomic Force Microscopy (AFM). We estimate the lifetime and the off rates of the formed intermediate.