PROTEIN-POLYMER COMPLEX, TGase SUBSTRATE-CONTAINING POLYMER, TGase SUBSTRATE-CONTAINING MONOMER, METHOD FOR PRODUCING PROTEIN-POLYMER COMPLEX, AND METHOD FOR IMPROVING PROTEIN FUNCTION AT SOLID-LIQUID INTERFACE OR IN VICINITY OF SOLID-LIQUID INTERFACE

Patent metadata

Field	Value
Publication number	US-2016251450-A1
Application number	US-201414392208-A
Country	US
Kind code	A1
Filing date	Jun 27, 2014
Priority date	Jun 28, 2013
Publication date	Sep 1, 2016
Grant date	—

Abstract

Provided is a protein-polymer complex which is capable of detecting a target with good sensitivity. Specifically provided is a protein-polymer complex comprising a polymer having a glutamine (Gln) residue or a primary amine on a side chain, wherein either a protein having a primary amine is bound to the glutamine (Gln) residue, or a protein having a glutamine (Gln) residue is bound to the primary amine.

First claim

1 . A protein-polymer complex wherein a protein having a primary amine is bound to a glutamine (Gln) residue in a polymer of monomers including an acrylate ester or a methacrylate ester having the glutamine (Gln) residue, represented by the following Formula (1):

wherein in Formula (1), R 1 represents a hydrogen atom or a protective group, R 2 represents a hydrogen atom or a methyl group, each of X and Y independently represents an alkylene group having a carbon number of 1 to 48 or an alkenylene group having a carbon number of 2 to 48, which may be substituted with an ethenylene group or —(C 2 H 4 O) n — or —(C 3 H 6 O) n —, wherein n represents the number of repeating units, and Z represents an alkyl group having a carbon number of 1 to 48, an alkoxy group having a carbon number of 1 to 48, an aryl group having a carbon number of 6 to 48, an aryloxy group having a carbon number of 6 to 48, an arylalkyl group having a carbon number of 7 to 48, or an arylalkyloxy group having a carbon number of 7 to 48, which may be substituted with a dinitrophenyl group or L-3,4-dihydroxyphenyl group, and wherein at least one of Y and Z may be independently substituted with an amino acid other than Lys.

2 . A TGase substrate-containing polymer of monomers including an acrylate ester or a methacrylate ester having a glutamine (Gln) residue, represented by the following Formula (1):

wherein in Formula (1), R 1 represents a hydrogen atom or a protective group, R 2 represents a hydrogen atom or a methyl group, each of X and Y independently represents an alkylene group having a carbon number of 1 to 48 or an alkenylene group having a carbon number of 2 to 48, which may be substituted with an ethenylene group or —(C 2 H 4 O) n — or —(C 3 H 6 O) n —, wherein n represents the number of repeating units, and Z represents an alkyl group having a carbon number of 1 to 48, an alkoxy group having a carbon number of 1 to 48, an aryl group having a carbon number of 6 to 48, an aryloxy group having a carbon number of 6 to 48, an arylalkyl group having a carbon number of 7 to 48, or an arylalkyloxy group having a carbon number of 7 to 48, which may be substituted with a dinitrophenyl group or L-3,4-dihydroxyphenyl group, and wherein at least one of Y and Z may be independently substituted with an amino acid other than Lys.

3 . A TGase substrate-containing monomer, which is an arcylate ester or a methacrylate ester having a glutamine (Gln) residue represented by following Formula (1), that can be used in production of a polymer having a glutamine (Gln) residue on a side chain:

wherein in Formula (1), R 1 represents a hydrogen atom or a protective group, R 2 represents a hydrogen atom or a methyl group, each of X and Y independently represents an alkylene group having a carbon number of 1 to 48 or an alkenylene group having a carbon number of 2 to 48, which may be substituted with an ethenylene group or —(C 2 H 4 O) n — or —(C 3 H 6 O) n —, wherein n represents the number of repeating units, and Z represents an alkyl group having a carbon number of 1 to 48, an alkoxy group having a carbon number of 1 to 48, an aryl group having a carbon number of 6 to 48, an aryloxy group having a carbon number of 6 to 48, an arylalkyl group having a carbon number of 7 to 48, or an arylalkyloxy group having a carbon number of 7 to 48, which may be substituted with a dinitrophenyl group or L-3,4-dihydroxyphenyl group, and wherein at least one of Y and Z may be independently substituted with an amino acid other than Lys.

4 . A method for producing a protein-polymer complex, the method comprising a protein binding step of using a transglutaminase (TGase) to bind a protein having a primary amine to a glutamine (Gln) residue of a polymer of monomers including an acrylate ester or a methacrylate ester having the glutamine (Gln) residue represented by following Formula (1):

wherein in Formula (1), R 1 represents a hydrogen atom or a protective group, R 2 represents a hydrogen atom or a methyl group, each of X and Y independently represents an alkylene group having a carbon number of 1 to 48 or an alkenylene group having a carbon number of 2 to 48, which may be substituted with an ethenylene group or —(C 2 H 4 O) n — or —(C 3 H 6 O) n —, wherein n represents the number of repeating units, and Z represents an alkyl group having a carbon number of 1 to 48, an alkoxy group having a carbon number of 1 to 48, an aryl group having a carbon number of 6 to 48, an aryloxy group having a carbon number of 6 to 48, an arylalkyl group having a carbon number of 7 to 48, or an arylalkyloxy group having a carbon number of 7 to 48, which may be substituted with a dinitrophenyl group or L-3,4-dihydroxyphenyl group, and wherein at least one of Y and Z may be independently substituted with an amino acid other than Lys.

5 . A method for detecting a protein, the method comprising binding, either directly or indirectly, a protein-polymer complex in which a protein having a primary amine is bound to a glutamine (Gln) residue in a polymer of monomers including an acrylate ester or a methacrylate ester having the glutamine (Gln) residue represented by following Formula (1), and a target protein that exists within a target material, binding a labeling molecule comprising a portion that binds specifically to the protein of the bound protein-polymer complex, and detecting the labeling molecule,

wherein in Formula (1), R 1 represents a hydrogen atom or a protective group, R 2 represents a hydrogen atom or a methyl group, each of X and Y independently represents an alkylene group having a carbon number of 1 to 48 or an alkenylene group having a carbon number of 2 to 48, which may be substituted with an ethenylene group or —(C 2 H 4 O) n — or —(C 1 H 6 O) n —, wherein n represents the number of repeating units, and Z represents an alkyl group having a carbon number of 1 to 48, an alkoxy group having a carbon number of 1 to 48, an aryl group having a carbon number of 6 to 48, an aryloxy group having a carbon number of 6 to 48, an arylalkyl group having a carbon number of 7 to 48, or an arylalkyloxy group having a carbon number of 7 to 48, which may be substituted with a dinitrophenyl group or L-3,4-dihydroxyphenyl group, and wherein at least one of Y and Z may be independently substituted with an amino acid other than Lys.

6 . An enzyme reaction method, wherein a protein-polymer complex in which a protein having a primary amine is bound to a glutamine (Gln) residue in a polymer of monomers including an acrylate ester or a methacrylate ester having the glutamine (Gln) residue represented by following Formula (1), and in which the protein is an enzyme, acts upon a substrate,

wherein in Formula (1), R 1 represents a hydrogen atom or a protective group, R 2 represents a hydrogen atom or a methyl group, each of X and Y independently represents an alkylene group having a carbon number of 1 to 48 or an alkenylene group having a carbon number of 2 to 48, which may be substituted with an ethenylene group or —(C 2 H 4 O) n — or —(C 3 H 6 O) n —, wherein n represents the number of repeating units, and Z represents an alkyl group having a carbon number of 1 to 48, an al

Assignees

• Univ Kyushu Nat Univ Corp
• Hitachi Aloka Medical Ltd

Inventors

• Kamiya Noriho
• Wakabayashi Rie
• Yahiro Kensuke
• Hayashi Kounosuke

Classifications

• C12N9/2437

  Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150) · CPC title

• C07K5/06034

  the side chain containing 2 to 4 carbon atoms · CPC title

• C07K14/00 Primary

  Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof · CPC title

• G01N33/68

  involving proteins, peptides or amino acids {(involving lipoproteins G01N33/92)} · CPC title

• G01N33/54306

  Solid-phase reaction mechanisms · CPC title