System and method for identification and characterization of transglutaminase species
In one aspect, the present disclosure provides a system and method for the identification and characterization of a transglutaminase. Further, the present disclosure provides transglutaminase enzymes for forming isopeptide bonds, methods of forming isopeptide bonds in the presence of transglutaminases, and substrate tags for use with transglutaminases.
1. A structure, a substrate tag attached to the structure, and a microbial transglutaminase bound to the substrate tag, wherein the substrate tag comprises an acyl-donor tag having at least 80% sequence identity to a peptide sequence consisting of RYRQR (SEQ ID NO:14), wherein the acyl-donor tag is a substrate for the microbial transglutaminase, wherein the structure is selected from a recombinant protein, a detectable label, and a chemically synthesized structure, and wherein the microbial transglutaminase has at least 80% sequence identity to the Kutzneria albida microbial transglutaminase (SEQ ID NO:6).
2. The structure of claim 1 , wherein the detectable label is selected from a biotin moiety, a fluorescent dye, a ruthenium label, a radiolabel, and a chemiluminescent label.
3. The structure of claim 1 , wherein the acyl-donor tag has the peptide sequence APRYRQRAA (SEQ ID NO:24), and wherein the microbial transglutaminase is further bound to an amine-donor tag.
4. The structure of claim 1 , wherein the recombinant microbial transglutaminase is the Kutzneria albida microbial transglutaminase (SEQ ID NO:6).
5. The structure of claim 1 , wherein the chemically synthesized structure comprises at least one of a protein, a peptide, an oligonucleotide, a carboxybenzyl group, and a polyethylene glycol (PEG) polymer.
6. The structure of claim 1 , wherein the microbial transglutaminase is further bound to an amine-donor tag.
7. The structure of claim 6 , wherein the amine-donor tag has a peptide sequence consisting of RYESK (SEQ ID NO:2).
8. The structure of claim 7 , wherein the amine-donor tag is attached to a second structure, wherein the second structure is selected from a second recombinant protein, a second detectable label, and a second chemically synthesized structure.
9. The structure of claim 8 , wherein the second detectable label is selected from a biotin moiety, a fluorescent dye, a ruthenium label, a radiolabel, and a chemiluminescent label.
10. The structure of claim 8 , wherein the second chemically synthesized structure comprises at least one of a protein, a peptide, an oligonucleotide, a carboxybenzyl group, and a polyethylene glycol (PEG) polymer.