System and method for identification and characterization of transglutaminase species

Patent metadata

Field	Value
Publication number	US-10788495-B2
Application number	US-201815974385-A
Country	US
Kind code	B2
Filing date	May 8, 2018
Priority date	Dec 19, 2014
Publication date	Sep 29, 2020
Grant date	Sep 29, 2020

Abstract

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.

First claim

What is claimed is:

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.

Assignees

• Roche Sequencing Solutions Inc

Inventors

• Albert Thomas
• Bergmann Frank
• Lyamichev Victor
• Patel Jigar
• Schraeml Michael
• Steffen Wojtek
• Streidl Thomas

Classifications

• C07K1/1075 Primary

  by covalent attachment of amino acids or peptide residues · CPC title

• C07K7/06 Primary

  having 5 to 11 amino acids · CPC title

• C12N9/1044

  Protein-glutamine gamma-glutamyltransferase (2.3.2.13), i.e. transglutaminase or factor XIII · CPC title

• C07K14/21

  from Pseudomonadaceae (F) · CPC title

• G01N33/573 Primary

  for enzymes or isoenzymes · CPC title