title: Assessing the Catalytic Activity of Transglutaminases in the Context of Autophagic Responses.
abstract: The human transglutaminases (TGases) are a widely distributed and peculiar group of enzymes that catalyze the posttranslational modification of proteins by the formation of isopeptide bonds. Tissue or type 2 transglutaminase (TG2) represents the most ubiquitous isoform belonging to TGases family. The vast array of biochemical functions catalyzed by TG2 distinguishes it from the other members of the TGase family. In the presence of high calcium levels TG2 catalyzes a vast array of protein posttranslational modifications, including protein-protein cross-linking, incorporation of primary amines into proteins, as well as glutamine deamination. In the last few years, it has become evident that TG2 is involved in the final maturation of autolysosomes. The TG2 regulation of autophagy occurs by its transamidating activity and its inhibition results in the intracellular increase of ubiquitinated protein aggregates. In this chapter, we describe the methods used in our laboratories to assess the catalytic activity of TG2 in the autophagic process.
authors.affiliation: University of Rome "Tor Vergata", Rome, Italy.
authors.forename: M
authors.initials: M
authors.lastname: D'Eletto
authors.affiliation: University of Rome "Tor Vergata", Rome, Italy.
authors.forename: M G
authors.initials: MG
authors.lastname: Farrace
authors.affiliation: University of Rome "Tor Vergata", Rome, Italy; National Institute for Infectious Diseases, IRCCS "Lazzaro Spallanzani", Rome, Italy. Electronic address: mauro.piacentini@uniroma2.it.
authors.forename: M
authors.initials: M
authors.lastname: Piacentini
authors.affiliation: University of Rome "Tor Vergata", Rome, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Rossin
mesh_terms: D000818:Animals; D001343:Autophagy / Q000502:physiology*; D019204:GTP-Binding Proteins / Q000378:metabolism; D006801:Humans; D051379:Mice; D008967:Molecular Biology / Q000379:methods*; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D016601:RNA-Binding Proteins / Q000378:metabolism; D011503:Transglutaminases / Q000032:analysis / Q000378:metabolism*
chemical_list: C118585:P62 protein, human; D016601:RNA-Binding Proteins; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins
keywords: Autophagy; Starvation; Transamidating activity; Transglutaminase type 2; p62