Protein labeling with cyanobenzothiazole conjugates

Patent metadata

Field	Value
Publication number	US-9459249-B2
Application number	US-38383209-A
Country	US
Kind code	B2
Filing date	Mar 26, 2009
Priority date	Mar 27, 2008
Publication date	Oct 4, 2016
Grant date	Oct 4, 2016

Abstract

The invention provides compounds and methods for site-specifically labeling proteins with cyanobenzothiazole derivatives of formula I. For example, the invention provides methods for labeling the N-terminus of a protein that terminates with a cysteine residue. The invention also provides methods for isolating an N-terminally labeled protein and methods for detecting an N-terminally labeled protein.

First claim

What is claimed is:

1. A compound of formula I:

wherein
Z is H, F, Cl, Br, I, CN, amino, alkylamino, dialkylamino, alkyl ester, carboxy, carboxylic acid salt, alkyl amide, phosphate, alkyl phosphonate, sulfate, alkyl sulfonate, nitro, or (C 1 -C 10 )alkyl optionally unsaturated and optionally substituted with amino, hydroxy, oxo (═O), nitro, thiol, or halo;
each R 1 is independently H, F, Cl, Br, I, CN, (C 1 -C 6 )alkyl, (C 1 -C 6 )alkoxy, or (C 1 -C 6 )alkylthio, wherein each alkyl, alkoxy, or alkylthio is optionally substituted with F, Cl, Br, I, amino, alkenyl, alkynyl, cycloalkyl, aryl, alkyl sulfonate, or CO 2 M wherein M is H, an organic cation, or an inorganic cation;
n is 0, 1, or 2;
Y is a linking group comprising (C 1 -C 16 )alkyl optionally substituted with one or more halo, oxo (═O), (C 1 -C 6 )alkyl, or (C 1 -C 6 )alkoxy, and optionally interrupted with one or more N(R 1 ), O, S, or —N—C(═O)— groups; or Y is optionally absent when X is N 3 ; and
X is a fluorescent dye, biotin, HisTag, chitin, a solid support, N 3 , H, or OH, provided that when X is H or OH, the compound of formula I comprises a radioactive moiety or an isotopic variant of any atom other than the carbon or nitrogen atom of the 2-nitrile moiety.

2. The compound of claim 1 wherein Z is H, F, nitro, or alkyl sulfonate, and R 1 is H or F.

3. The compound of claim 2 wherein Y is —(C 1 -C 6 )alkyl-, —O—(C 1 -C 6 )alkyl-, —O—(C 1 -C 6 )alkyl-O—, —O—(C 1 -C 6 )alkyl-NH—, —O—(C 1 -C 6 )alkyl-(CO)NH—, —NH—(C 1 -C 6 )alkyl-NH—, —NH—(CO)(C 1 -C 6 )alkyl-NH—, —NH—(CO)(C 1 -C 6 )alkyl-(CO)—NH—, or —O—(C 1 -C 6 )alkyl-(CO)NH—(C 1 -C 6 )alkyl-.

4. The compound of claim 3 wherein X is N 3 , a fluorescent dye, biotin, HisTag, chitin, or a solid support.

5. The compound of claim 4 , wherein X is N 3 ,

6. The compound of claim 1 wherein the compound of formula I is:

Assignees

• Anderson Jessica
• Meisenheimer Poncho
• Shultz John
• Promega Corp

Inventors

• Anderson Jessica
• Meisenheimer Poncho
• Shultz John

Classifications

• G01N33/533 Primary

  with fluorescent label · CPC title

• C09B11/24

  Phthaleins containing amino groups {; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes} · CPC title

• C09B23/083

  five >CH- groups · CPC title

• C07D493/10

  Spiro-condensed systems · CPC title

• C09B11/08

  Phthaleins; {Phenolphthaleins; Fluorescein} · CPC title