title: Identification of a highly reactive substrate peptide for transglutaminase 6 and its use in detecting transglutaminase activity in the skin epidermis.
abstract: Mammalian transglutaminases (TGs) are a family of enzymes that catalyze the formation of covalent crosslinks between glutamine and lysine residues in proteins. These catalytic reactions play roles in several essential biological processes, including blood coagulation, skin formation, and stabilization of the extracellular matrix. Among the members of this family, factor XIII and TGs 1-5 have been characterized well, but very little is known about the novel members TG6 and TG7. Recently, however, autoantibodies against TG6 were found in a patient with gluten ataxia, a disease caused by enzymatically modified gluten-derived peptides in neuronal cells. To characterize the possible physiological functions of TG6, in this study we screened a phage-displayed random peptide library to find highly reactive glutamine donor substrate peptides. From several candidate peptides, one sequence, designated Y25, appeared to have the highest reactivity. The Y25 sequence also has apparent isozyme specificity when evaluated by incorporation of the labeled glutamine acceptor substrate as a fusion protein with glutathione-S-transferase. Also, the sequence retained high reactivity as well as the isozyme specificity in the peptide form. Analyses with the biotin-labeled and fluorescence-labeled peptides showed TG6 to be an active enzyme and react to specific substrates in the skin, which is consistent with the results of the expression pattern of its transcripts.
authors: Graduate School of Pharmaceutical Sciences, Nagoya University, Nagoya, Japan.
authors: Mina
authors: M
authors: Fukui
authors: Katsuma
authors: K
authors: Kuramoto
authors: Risa
authors: R
authors: Yamasaki
authors: Yoshitaka
authors: Y
authors: Shimizu
authors: Miho
authors: M
authors: Itoh
authors: Tadafumi
authors: T
authors: Kawamoto
authors: Kiyotaka
authors: K
authors: Hitomi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn / Q000378:metabolism; D001710:Biotin / Q000378:metabolism; D004789:Enzyme Activation; D057075:Enzyme Assays; D000078404:Epidermal Cells; D004817:Epidermis / Q000201:enzymology*; D015971:Gene Expression Regulation, Enzymologic; D005973:Glutamine / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D051379:Mice; D019151:Peptide Library; D010455:Peptides / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012867:Skin / Q000166:cytology / Q000201:enzymology*; D013379:Substrate Specificity; D011503:Transglutaminases / Q000235:genetics / Q000378:metabolism*
chemical_list: D007527:Isoenzymes; D019151:Peptide Library; D010455:Peptides; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D005973:Glutamine; D001710:Biotin; C532876:TGM6 protein, human; D011503:Transglutaminases; D005982:Glutathione Transferase