Astexin peptides

Patent metadata

Field	Value
Publication number	US-10927152-B2
Application number	US-201816057292-A
Country	US
Kind code	B2
Filing date	Aug 7, 2018
Priority date	Aug 31, 2012
Publication date	Feb 23, 2021
Grant date	Feb 23, 2021

Abstract

Provided are astexin-1, astexin-2 and astexin-3 lasso peptides, which are based on sequences identified in Asticcacaulis excentricus , and methods of making and using same.
Astexin-1 is highly polar, in contrast to many lasso peptides that are primarily hydrophobic, and has modest antimicrobial activity against Caulobacter crescentus , a bacterium related to Asticcacaulis excentricus . The solution structure of astexin-1 was determined, revealing a unique topology that is stabilized by hydrogen bonding between segments of the peptide. Astexins-2 and -3 are intracellular lasso peptides.

First claim

What is claimed:

1. A substantially purified Astexin-1 peptide consisting of the amino acid sequence GLSQGVEPDIGQTYFEESRINQD (SEQ ID NO:3);
wherein the glycine residue (G) at position 1 is covalently bound to the aspartic acid residue (D) at position 9, thereby generating a lassoed peptide, and
wherein the C-terminal amino acid of the peptide comprises a protecting group.

2. An Astexin-1 fusion protein comprising the amino acid sequence GLSQGVEPDIGQTYFEESRINQD (SEQ ID NO:3) and a second protein moiety, wherein the second protein moiety is linked to the C-terminus of SEQ ID NO:3.

3. The Astexin-1 fusion protein of claim 2 , wherein the glycine residue (G) at position 1 of SEQ ID NO:3 is covalently bound to the aspartic acid residue (D) at position 9 of SEQ ID NO:3, thereby generating a lassoed fusion protein.

4. A non-naturally occurring polynucleotide sequence encoding the peptide of claim 1 , wherein the non-naturally occurring polynucleotide sequence includes at least one codon that differs from a naturally occurring polynucleotide sequence.

5. The non-naturally occurring polynucleotide sequence according to claim 4 , wherein the non-naturally occurring polynucleotide sequence includes a cis acting regulatory element.

6. The non-naturally occurring polynucleotide sequence according to claim 4 , further comprising a selectable marker, an origin of replication, or both.

7. A non-naturally occurring polynucleotide sequence encoding the peptide of claim 2 .

Assignees

• Univ Princeton

Inventors

• Link A James
• Maksimov Mikhail O

Classifications

• A01N63/50

  Isolated enzymes; Isolated proteins (peptides A01N37/46) · CPC title

• C02F1/286

  using natural organic sorbents or derivatives thereof · CPC title

• C07K14/195 Primary

  from bacteria · CPC title

• A62D3/33

  by chemical fixing the harmful substance, e.g. by chelation or complexation · CPC title

• C02F2101/20

  Heavy metals or heavy metal compounds · CPC title