Transglutaminase amine substrates for photochemical labeling and cleavable cross-linking of proteins.
A new procedure for the photochemical labeling of peptides and for the production of cleavable cross-links between protein molecules is given. This method is mediated through the catalytic action of the enzyme guinea pig liver transglutaminase. Each of the labeling and cross-linking reagents described here is an amine substrate for transglutaminases and, because of the narrow specificity of these enzymes, is introduced covalently only at the gamma-carboxamide group of available peptide-bound glutamine residues. Cross-linking results either solely through the action of the enzyme in the case of a diamine substrate, or by subsequent photolysis in the case of photosensitive amine substrates. Cleavable bonds in several of the substrates are disulfide or vicinal hydroxyl groups. The validity of the procedure is demonstrated by the preparation of photosensitive derivatives of substance P and glucagon 1-6 and in the cleavable covalent cross-linking of guanidinated beta-casein.
D000345:Affinity Labels; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D003432:Cross-Linking Reagents; D006168:Guinea Pigs; D008099:Liver / Q000201:enzymology; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D010446:Peptide Fragments / Q000032:analysis; D010777:Photochemistry; D013053:Spectrophotometry; D013379:Substrate Specificity; D005723:gamma-Glutamyltransferase / Q000378:metabolism*
D000345:Affinity Labels; D000596:Amino Acids; D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D010446:Peptide Fragments; D005723:gamma-Glutamyltransferase