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Methods and compositions for protein labeling using lipoic acid ligases
US9284541B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9284541-B2 |
| Application number | US-201113267761-A |
| Country | US |
| Kind code | B2 |
| Filing date | Oct 6, 2011 |
| Priority date | Nov 9, 2007 |
| Publication date | Mar 15, 2016 |
| Grant date | Mar 15, 2016 |
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- Title
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Abstract
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The present disclosure provides compositions and methods of use thereof for labeling peptide and proteins in vitro or in vivo. The methods described herein employ lipoic acid ligase or mutants thereof, and lipoic acid analogs (e.g., lipoic acid analogs comprising a resorufin moiety) recognized by lipoic acid ligase and lipoic acid ligase mutants. Also provided herein is a method of imaging protein-protein interaction via a reaction mediated by lipoic acid ligase.
First claim
Opening claim text (preview).
What is claimed is: 1. An isolated lipoic acid ligase mutant that uses resorufin as a substrate, the mutant comprising an amino acid sequence that is at least 85% identical to SEQ ID NO:11 and includes mutations at residues corresponding to E20 and F147 in SEQ ID NO:11. 2. The lipoic acid ligase mutant of claim 1 , wherein the mutant further comprises a mutation at the residue corresponding to H149 in SEQ ID NO:11. 3. The lipoic acid ligase mutant of claim 1 , wherein the amino acid sequence is at least 95% identical to SEQ ID NO:11. 4. The lipoic acid ligase mutant of claim 2 , wherein the mutant includes amino acid residue substitutions corresponding to E20A, F147A, and H149G. 5. The lipoic acid ligase mutant of claim 2 , wherein the amino acid sequence of the mutant is otherwise identical to SEQ ID NO:11 except for the E20A, F147A, and H149G substitutions.
Assignees
Inventors
Classifications
- C12N9/1241Primary
Nucleotidyltransferases (2.7.7) · CPC title
Lipoate--protein ligase (2.7.7.63) · CPC title
Ligases (6) · CPC title
Mutagenizing nucleic acids · CPC title
Production of labelled immunochemicals · CPC title
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- What does patent US9284541B2 cover?
- The present disclosure provides compositions and methods of use thereof for labeling peptide and proteins in vitro or in vivo. The methods described herein employ lipoic acid ligase or mutants thereof, and lipoic acid analogs (e.g., lipoic acid analogs comprising a resorufin moiety) recognized by lipoic acid ligase and lipoic acid ligase mutants. Also provided herein is a method of imaging prot…
- Who is the assignee on this patent?
- Ting Alice Y, Baker David, Richter Florian, and 4 more
- What technology area does this patent fall under?
- Primary CPC classification C12N9/1241. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Mar 15 2016 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 8 related publications on this page (citations in our corpus or others sharing the same primary CPC).