title: Comparison of substrate specificities of transglutaminases using synthetic peptides as acyl donors.
abstract: Transglutaminase (TGase) is an enzyme that catalyzes acyl transfer reactions between primary amines and Gln residues in proteins or peptides. Substrate specificities of TGase, Ca2+-independent microbial transglutaminase (MTGase), and Ca2+-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.
authors.affiliation: Food Research and Development Laboratories, Ajinomoto Co., Inc., Kawasaki-shi, Japan. tomoko_ootsuka@ajinomoto.com
authors.forename: T
authors.initials: T
authors.lastname: Ohtsuka
authors.forename: M
authors.initials: M
authors.lastname: Ota
authors.forename: N
authors.initials: N
authors.lastname: Nio
authors.forename: M
authors.initials: M
authors.lastname: Motoki
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D005973:Glutamine / Q000737:chemistry / Q000378:metabolism; D006168:Guinea Pigs; D007930:Leucine / Q000737:chemistry / Q000378:metabolism; D008099:Liver / Q000201:enzymology; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D013045:Species Specificity; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D011503:Transglutaminases / Q000378:metabolism*
chemical_list: D010455:Peptides; D005973:Glutamine; D011503:Transglutaminases; D007930:Leucine