Transglutaminase, assay

Slaughter, T.F., Achyuthan, K.E., Lai, T.-S., and Greenberg, C.S. (1992) A microtiter plate transglutaminase assay utilizing 5-(biotinamido)pentylamine as substrate. Anal. Biochem. 205, 1-6. 

Sulkanen S, Halttunen T, Laurila K, Kolho KL, Korponay-Szabo IR, Samesto A, et al. Tissue transglutaminase autoantibody enzyme-linked immunosorbent assay in detecting celiac disease. Gastroenterology 1998 115 1322-1328. 

Seissler J, Boms S, Wohlrab U, Morgenthaler NG, Mothes T, Boehm BO, et al. Antibodies to human recombinant tissue transglutaminase measured by radioligand assay Evidence for high diagnostic sensitivity for celiac disease. Horm Metab Res 1999 31 375-379. 

Sblattero D, Berti I, Trevisiol C, Marzari R, Tommasini A, Bradbury A, et al. Human recombinant tissue transglutaminase ELISA An innovative diagnostic assay for celiac disease. Am J Gastroenterol 2000 95 1253-1257. 

Shamir R, Eliakim R, Lahat N, Sobel E, Lemer A. ELISA of anti-endomysial antibodies in the diagnosis of celiac disease Comparison with immunofluorescence assay of anti-endomysial antibodies and tissue transglutaminase antibodies. Isr Med Assoc J 2002 4 594-596. 

Mamone G, Ferranti P, Melck D, Tafuro F, Longobardo L, Chianese L, et al. Susceptibility to transglutaminase of gliadin peptides predicted by a mass spectrometry-based assay. FEBS Lett 2004 562 177-182. 

Application and Principle This procedure is used to determine transglutaminase activity in preparations derived from Streptoverticillium mobaraense var. The assay is based on the enzymatic formation of a glutamic acid y-hydroxamate in a glutaminyl residue in the substrate peptide with another substrate, hydroxylamine. The amount of the glutamic acid y-hydroxamate formed as a red complex with ferric ion in acidic conditions at 37° is measured spectrophotometrically. 

Calculation One unit of enzyme activity is defined as the amount of enzyme that catalyzes the transglutamination of 1 pimol of substrate into product in 1 min under the conditions of the assay. The specific activity of Transglutaminase is defined as... 

Transglutaminase catalyzes cross-linking between peptide chains in fibrin clots, the cornihed envelope of epidermal cells, and the vaginal plug from seminal plasma in rodents. Glutaminyl residues serve as acyl donors to peptide-bound lysine. This assay makes use of a small synthetic peptide, benzyloxycarbonyl-L-Gln-Gly, and fluorescent amine monodansylcadaverine as substrates. 

Gnaccarini, C. Ben-Tahar, W Lubell, W. D. Pelletier, J. N. Keillor, J. W. Fluorometric assay for tissue transglutaminase-mediated transamidation activity. Bioorg. Med. Chem. 2009,17, 6354-6359. 

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