Abstract
The identification of both acyl donor and acceptor substrates, as well as the detection of e( γ -glutamyl)lysine crosslinks and ( γ-glutamyl) amine derivatives, are some of the most important aspects in the field of the transglutaminase (TGase)-mediated reactions1–5. These reactions have been revealed so far using both direct and indirect methodologies. The main direct method is based upon chromatographic isolation and identification of the isopeptide e( γ -glutamyl)lysine or of ( γ -glutamyl)amine derivatives after exhaustive proteolytic digestion of the reaction products. On the other hand, the detection of either polymers or radioactive amine-protein adducts by SDS-PAGE, together with the protein crosslinking inhibition by small molecular weight amines, represent the most commonly used indirect methodologies. However, both procedures suffer from severe limitations since a single analytical system does not yield unambiguous results.
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