Methods for ligation and uses thereof

• US 8,940,501 B2
• Filed: 02/01/2010
• Issued: 01/27/2015
• Est. Priority Date: 01/30/2009
• Status: Active Grant

• Current Assignee: Whitehead Institute for Biomedical Research
• Original Assignee: Whitehead Institute for Biomedical Research
• Inventors: Ploegh, Hidde L., Antos, John M., Guimaraes, Carla, Popp, Maximilian Wei-Lin
• Primary Examiner(s): Skowronek, Karlheinz R
• Assistant Examiner(s): NEWMAN, MINDY B
• Application Number:
  US13/145,644
• Publication Number:
  US 20110321183A1
• Time in Patent Office: 1,821 Days
• Field of Search: None
• US Class Current: 435/68.1
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• CPC Class Codes: C12N 9/48 acting on peptide bonds (3.4)
• CPC Class Codes: C12N 9/80 acting on amide bonds in linear amides (3.5.1)

Abstract

The present invention relates to methods for ligation. The invention provides novel reagents and methods for ligating an acyl donor compound with an acyl acceptor compound. Provided acyl donor compounds comprise a transamidase recognition sequence that allows ligation with a nucleophilic acyl acceptor in the presence of transamidase. The invention further provides kits comprising acyl donor compounds and optionally comprising other reagents for ligation.

Claims

1. A method of ligation comprising the step of contacting an acyl donor compound of formula:

2. The method of claim 1, wherein X is —

O—

.

3. The method of claim 1, wherein the recognition sequence is LPXT, wherein the X of the recognition sequence is D, E, A, N, Q, K, or R.

4. The method of claim 1, wherein the transamidase is sortase A.

5. The method of ligation of claim 1 comprising the step of contacting an acyl donor compound of formula:

6. A method of ligation comprising the step of contacting an acyl donor compound of formula:

7. The method of claim 6, wherein X is —

O—

.

8. The method of claim 6, wherein the X of the recognition sequence is D, E, A, N, Q, K, or R.

9. The method of claim 6, wherein the transamidase is sortase A.

10. The method of claim 6, wherein the transamidase is sortase B.

11. The method of claim 6 comprising the step of contacting an acyl donor compound of formula:

12. A method of ligation comprising the step of contacting an acyl donor compound of formula:

13. The method of claim 12, wherein X is —

O—

.

14. The method of claim 12, wherein the transamidase is sortase A.

15. The method of claim 12, wherein the transamidase is sortase B.

16. The method of claim 12 comprising the step of contacting an acyl donor compound of formula:

17. A method of ligation comprising the step of contacting an acyl donor compound of formula:

18. The method of claim 17, wherein X is —

O—

.

19. The method of claim 17, wherein the transamidase is sortase A.

20. The method of claim 17, wherein the transamidase is sortase B.

21. The method of claim 17 comprising the step of contacting an acyl donor compound of formula: