Astexin peptides

Patent metadata

Field	Value
Publication number	US-11732013-B2
Application number	US-202017128632-A
Country	US
Kind code	B2
Filing date	Dec 21, 2020
Priority date	Aug 31, 2012
Publication date	Aug 22, 2023
Grant date	Aug 22, 2023

Abstract

Provided are astexin-1, astexin-2 and astexin-3 lasso peptides, which are based on sequences identified in Asticaccaulis 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 Asticaccaulis 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 is:

1. A substantially purified Astexin-3 peptide comprising the amino acid sequence GPTPMVGLDSVSGQYWDQHAPLAD (SEQ ID NO:7); 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.

2. The peptide of claim 1 , wherein the peptide is less than 50 amino acids.

3. The peptide of claim 1 , wherein the peptide is less than 35 amino acids.

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. An Astexin-3 fusion protein comprising the amino acid sequence GPTPMVGLDSVSGQYWDQHAPLAD (SEQ ID NO:7) and a second protein moiety, wherein the N-terminus of the second protein moiety is linked via a peptide bond to the C-terminus of SEQ ID NO:7.

8. The Astexin-3 fusion protein of claim 7 , wherein the glycine residue (G) at position 1 of SEQ ID NO:7 is covalently bound to the aspartic acid residue (D) at position 9 of SEQ ID NO:7, thereby generating a lassoed fusion protein.

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

10. A substantially purified Astexin-2 peptide comprising the amino acid sequence GLTQIQALDDSVSGQFRDQLGLSAD (SEQ ID NO:5) 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.

11. The peptide of claim 10 , wherein the peptide is less than 50 amino acids.

12. The peptide of claim 10 , wherein the peptide is less than 35 amino acids.

13. A non-naturally occurring polynucleotide sequence encoding the peptide of claim 10 , wherein the non-naturally occurring polynucleotide sequence includes at least one codon that differs from a naturally occurring polynucleotide sequence.

14. The non-naturally occurring polynucleotide sequence according to claim 13 , wherein the non-naturally occurring polynucleotide sequence includes a cis acting regulatory element.

15. The non-naturally occurring polynucleotide sequence according to claim 13 , further comprising a selectable marker, an origin of replication, or both.

16. An Astexin-2 fusion protein comprising the amino acid sequence GLTQIQALDDSVSGQFRDQLGLSAD (SEQ ID NO:5) and a second protein moiety, wherein the N-terminus of the second protein moiety is linked via a peptide bond to the C-terminus of SEQ ID NO:5.

17. The Astexin-2 fusion protein of claim 16 , wherein the glycine residue (G) at position 1 of SEQ ID NO:5 is covalently bound to the aspartic acid residue (D) at position 9 of SEQ ID NO:5, thereby generating a lassoed fusion protein.

18. A non-naturally occurring polynucleotide sequence encoding the peptide of claim 16 .

Assignees

• Univ Princeton

Inventors

• Link A James
• Maksimov Mikhail O

Classifications

• C07K14/195 Primary

  from bacteria · CPC title

• A01N63/50

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

• A62D3/33

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

• C02F1/286

  using natural organic sorbents or derivatives thereof · CPC title

• A62D2101/43

  containing heavy metals, in the bonded or free state · CPC title