Uranium-chelating peptides and uses thereof

Patent metadata

Field	Value
Publication number	US-9321849-B2
Application number	US-97978710-A
Country	US
Kind code	B2
Filing date	Dec 28, 2010
Priority date	Jul 4, 2003
Publication date	Apr 26, 2016
Grant date	Apr 26, 2016

Abstract

The invention concerns uranium-chelating peptides as well as their uses for decontaminating soils and water, and for detecting and treating people contaminated by uranium. Said peptides have a helix-loop-helix type structure comprising the sequence of a calmodulin loop including at least one mutation of neutral residues selected from the group consisting of S, T, C, H, Y, N and Q, of one, two or three residues of at least one of the four calmodulin calcium binding sites: site I: residues selected among D20, D22 and D24 residues; site II: residues selected among D56, D58 and N60 residues; site III: residues selected among D93, D95 and N97 residues; site IV: residues selected among D129, D131 and D133 residues; said positions being indicated with reference to the human calmodulin sequence.

First claim

The invention claimed is:

1. A peptide comprising one of the sequences SEQ ID Nos. 4-7 or SEQ ID Nos. 9-12.

2. The peptide as claimed in claim 1 , wherein cysteines at positions 13 and 29 of said sequences are connected via a disulfide bridge.

3. The peptide as claimed in claim 1 , wherein it is conjugated to at least one fluorophore.

4. The peptide as claimed in claim 3 , wherein it is conjugated to two different fluorophores.

5. The peptide as claimed in claim 3 wherein said fluorophore is a fluorescent protein selected from: EBFP, ECFP, EYFP, EGFP, DsRed, CopGFP and PhiYFP.

6. The peptide as claimed in claim 3 wherein said fluorophore is selected from dansyl, coumarin, fluorescein and Alexa derivatives.

7. The peptide as claimed in claim 1 , wherein it is associated with at least one molecule that allows targeting to the kidney and/or to the bone.

8. The peptide as claimed in claim 1 , wherein it is associated with a molecule that promotes its excretion in vivo.

9. A polypeptide comprising the concatenation of at least two identical or at least two different peptides as claimed in claim 1 .

10. A peptide composition comprising at least one polypeptide as claimed in claim 9 and at least one suitable vehicle.

11. A fusion protein consisting of the in-frame fusion of the sequence of at least one peptide as claimed in claim 1 , with the sequence of protein selected from the group consisting of calmodulin, chameleon proteins, and proteins having a helix-loop-helix type motif, capable of binding calcium.

12. The fusion protein as claimed in claim 11 wherein it is conjugated to at least one fluorophore as defined in claim 5 .

13. The fusion protein as claimed in claim 12 , wherein one of the ends of said protein is coupled to a fluorescence donor, and the other is coupled to a fluorescence acceptor.

14. The fusion protein as claimed in claim 13 , wherein it comprises, at one of its ends, the sequence of EBFP or ECFP and, at the other end, the sequence of EGFP or of EYFP.

15. A kit for detecting a contamination with uranium comprising at least one of: a peptide as claimed in claim 1 , a polypeptide derived from said peptide, a peptide composition comprising said peptide, a fusion protein derived from said peptide, an antibody that binds selectively to said peptide, or modified prokaryotic or eukaryotic cells modified with a regulatory system that includes said peptide as a regulator or repressor of a gene encoding a bioluminescent protein.

16. The fusion protein as claimed in claim 11 wherein it is conjugated to at least one fluorophore selected from the group consisting of dansyl, coumarin, fluorescein and Alexa derivatives.

Assignees

• Vita Claudio
• Sauvage Vita Mireille
• Vita Fabio
• Vita Elena
• Le Clainche Loïc
• Monjardet Véronique
• Commissariat Energie Atomique

Inventors

• Vita Claudio
• Le Clainche Loïc
• Monjardet Véronique

Classifications

• G01N2333/4727

  Calcium binding proteins, e.g. calmodulin · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• C07K14/4728

  Calcium binding proteins, e.g. calmodulin · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• C07K19/00 Primary

  Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} · CPC title