Labeling of epsilon-lysine crosslinking sites in proteins with peptide substrates of factor XIIIa and transglutaminase
| Autor: | Pauline T. Velasco, K. N. Parameswaran, James H. Wilson, Laszlo Lorand |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1990 |
| Předmět: |
Tissue transglutaminase
Lysine Molecular Sequence Data Peptide Substrate Specificity chemistry.chemical_compound Cadaverine Lens Crystalline Peptide synthesis Animals Amino Acid Sequence Peptide sequence chemistry.chemical_classification Fibrin Multidisciplinary Binding Sites Transglutaminases biology Crystallins Glutamine Kinetics Cross-Linking Reagents Biochemistry chemistry biology.protein Indicators and Reagents Pyroglutamic acid Factor XIIIa Rabbits Oligopeptides Research Article |
| Popis: | Peptides patterned on the N-terminal sequence of fibronectin were synthesized and tested for amine acceptor qualities in reactions with dansylcadaverine catalyzed either by coagulation factor XIIIa or intracellular transglutaminase (protein-glutamine:amine gamma-glutamyltransferase, EC 2.3.2.13). On the basis of inverse half-saturations of the enzymes, the order of acceptor substrate affinity for factor XIIIa was pEAQQIV much greater than Boc-AQQIV greater than Boc-QQIV, and for transglutaminase, Boc-QQIV greater than Boc-AQQIV greater than pEAQQIV (amino acid residues are shown in one-letter code; pE, pyroglutamic acid; Boc, tert-butyloxycarbonyl). Sequence analysis of dansylcadaverine-substituted pEAQQIV indicated that the first of the two adjacent glutamine residues was the target of enzymatic modification. Boc-QIV showed no substrate activity with either enzyme. Crosslinking of crystallins in Ca2(+)-treated rabbit lens homogenate was readily inhibited by Boc-QQIV, Boc-AQQIV, and pEAQQIV, as was the formation of alpha-chain polymers in human fibrin by pEAQQIV in the presence of human factor XIIIa. SDS/PAGE analysis suggested that the inhibitory peptides selectively blocked the electron donor functionalities in these enzymatic crosslinking reactions. |
| Databáze: | OpenAIRE |
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