Comparison of substrate specificities of transglutaminases using synthetic peptides as acyl donors

• Published in: Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2000-12, Vol.64 (12), p.2608-2613
• Main Authors: Ohtsuka, T. (Ajinomoto Co. Inc., Kawasaki, Kanagawa (Japan). Central Research Labs.), Ota, M, Nio, N, Motoki, M
• Format: Article
• Language: English
• Subjects: acyl donor; Amino Acid Substitution; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Glutamine - chemistry; Glutamine - metabolism; Guinea Pigs; Leucine - chemistry; Leucine - metabolism; Liver - enzymology; PEPTIDES; Peptides - chemical synthesis; Peptides - chemistry; Peptides - metabolism; Species Specificity; Structure-Activity Relationship; Substrate Specificity; synthetic peptide; TRANSFERASES; transglutaminase; Transglutaminases - metabolism; ε-(γ-glutamyl)lysine bond
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.64.2608

Transglutaminase (TGase) is an enzyme that catalyzes acyl transfer reactions between primary amines and Gln residues in proteins or peptides. Substrate specificities of TGase, Ca 2+ -independent microbial transglutaminase (MTGase), and Ca 2+ -dependent tissue type transglutaminase from guinea pig liver (GTGase) and fish, Red sea bream (Pagrus major), liver (FTGase), for acyl donors were investigated using synthetic peptides containing Gln residues and Gln analogues with different lengths of side chain. MTGase dose not recognize the Gln analogues as a substrate and has strict substrate specificities toward L-Gln. Substrate peptides with a variety of sequences around the Gln residue, GXXQXXG (X=G, A, S, L, V, F, Y, R, N, E, L) were synthesized and used as acyl donors. As an acyl acceptor, the fluorescent reagent monodancyl cadaverine was used and the reactions analyzed with RP-HPLC. Substitution of the C-terminal of a Gln residue with a hydrophobic amino acid accelerated the reaction by GTGase and FTGase. N-terminal substitution of Gln residues had similar effects on the reaction by MTGase.