Transglutaminases: purification and activity assays

Abstract

Transglutaminases (TGases) are a widely distributed family of proteins found in many tissues and body fluids of vertebrates. To date the following types have been distinguished: secretory, tissue, epidermal, keratinocyte, and hemocyte TGase as well as factor XIIIa and erythrocyte hand 4.2 TGases are difficult to isolate, as they tend to form irreversible aggregates under native conditions. In this review, the isolation procedures for the different types of TGases are summarized. The most common chromatographic separation methods used for TGase purification are size-exclusion and ion-exchange chromatography. Additionally, other chromatographic methods (hydrophobic-interaction, affinity, adsorption chromatography) and electrophoretic techniques [preparative isoelectric focusing, sodium dodecyl sulphate polyacrylamide gel electrophoresis and zone electrophoresis] are described. Based on the enzymatic function of TGases (cross-linking of a primary amine and peptide bound glutamine), several established activity assays are described.