Metal Abstraction Peptide and Uses Thereof
AssigneesEchogen, Inc., University of Kansas
InventorsJennifer Ann Stowell Laurence, Mary Elizabeth Krause, Timothy A. Jackson, George Laurence
Life of this patent
Estimating the term…
Abstract
Compositions comprising certain tripeptides which are capable of binding a metal in a square planar orientation, a square pyramidal orientation, or both, are disclosed in processes for modulating tagged peptides or proteins. Such compositions and processes may be used for site-specific chiral inversion of amino acids, site-specific peptide cleavage, and protein purification, among other uses.
Claims48
A method of performing a site-specific chiral inversion of a tripeptide comprising complexing a metal ion with a tripeptide having a sequence LXLC1LC2 to form a metal ion-DXLC1DC2 complex,
- wherein X is any amino acid such that the tripeptide and metal ion form a complex having a square planar orientation or square pyramidal orientation or both, and wherein C1 and C2 are the same or different;
- and wherein C1 is chosen from a cysteine and a cysteine-like nonnatural amino acid;
- and wherein C2 is chosen from a cysteine, a cysteine-like nonnatural amino acid, a histidine, and a histidine-like nonnatural amino acid.
(canceled)
(canceled)
(canceled)
(canceled)
An anti-oxidant composition comprising a metal ion complexed with a tripeptide having the sequence LXLC1LC2 to form a metal ion-DXLC1DC2 complex;
- wherein X is any amino acid such that the tripeptide and metal ion form a complex having a square planar orientation or square pyramidal orientation or both, and wherein C1 and C2 are the same or different;
- and wherein C1 is chosen from a cysteine and a cysteine-like nonnatural amino acid;
- and wherein C2 is chosen from a cysteine, a cysteine-like nonnatural amino acid, a histidine, and a histidine-like nonnatural amino acid.
A method of performing a site-specific cleavage of a peptide comprising:
- providing a peptide with a sequence XC1C2, wherein X is any amino acid such that the peptide and metal ion form a complex having a square planar orientation or square pyramidal orientation or both, and wherein CI and C2 are the same or different;
- and wherein C1 is chosen from a cysteine and a cysteine-like nonnatural amino acid;
- and wherein C2 is chosen from a cysteine, a cysteine-like nonnatural amino acid, a histidine, and a histidine-like nonnatural amino acid;
- and
- combining a partial chelator with a metal capable of reacting with the sequence XC1C2;
- and
- allowing cleavage of the peptide within or at a terminal portion of the sequence XC1C2.
(canceled)
(canceled)
A kit comprising a peptide having a sequence comprising XC1C2, wherein X is any amino acid, C1 and C2 are the same or different, C1 is chosen from a cysteine and a cysteine-like nonnatural amino acid, and C2 is chosen from a cysteine, a cysteine-like nonnatural amino acid, a histidine, and a histidine-like nonnatural amino acid;
- and a partial chelator with a metal capable of reacting with the sequence XC1C2.
A method of protein purification comprising;
- a) binding a first composition comprising a target protein associated with a linker and an affinity tag to an affinity material comprising a ligand capable of capturing the first composition;
- b) providing a metal-containing cleavage agent capable of detaching the target protein from the first composition such that the detached target protein is devoid of any supplementary amino acids originating from the linker or the affinity tag;
- and
- c) separating the target protein from the first composition.
A composition for purifying a protein comprising;
- a) an affinity tag, wherein the affinity tag is capable of binding to a ligand;
- and
- b) a linker associated with the affinity tag, wherein the linker is covalently attached to a target protein;
- wherein the entirety of the affinity tag and the linker are capable of being detached from the target protein by the addition of a cleavage agent comprising a metal ion.
(canceled)
(canceled)
Dependencies are inferred from the claim text.