Synthesis and evaluation of resins bearing substrate-like inhibitor functions for capturing copper amine oxidases

摘要

In order to make new tools available for investigating interactions between enzyme and substrate-like inhibitors in copper amine oxidases (CAOs), a class of enzymes that controls important cellular processes such as the crosslinking of elastin and collagen, cell proliferation and the regulation of intracellular polyamines, starting from a previous work we synthesized the poly(N,N-dimethylacrylamide)-based resin R2 and the new TentaGel resins T3 and T4 obtained by immobilizing onto commercial TentaGel S-Br properly designed CAO substrate-like inhibitor moieties. The anchoring of CAO substrate-like inhibitor moieties was carried out through stable ether bonds onto commercial TentaGel S-Br which contains bromomethyl groups susceptible to nucleophilic substitution reactions.We resorted to polyacrylamide gel electrophoresis (PAGE) experiments to ascertain the capability of the prepared resins to capture plasma amine oxidase (PAO) and diamine oxidase (DAO), members of the CAO family. The poly(N,N-dimethylacrylamide)-based resin R2 resulted able of blocking PAO and DAO being the first insoluble polymeric materials able to capture enzymes of the CAO family.