| First Author | Witte MD | Year | 2012 |
| Journal | Proc Natl Acad Sci U S A | Volume | 109 |
| Issue | 30 | Pages | 11993-8 |
| PubMed ID | 22778432 | Mgi Jnum | J:186476 |
| Mgi Id | MGI:5432421 | Doi | 10.1073/pnas.1205427109 |
| Citation | Witte MD, et al. (2012) Preparation of unnatural N-to-N and C-to-C protein fusions. Proc Natl Acad Sci U S A 109(30):11993-8 |
| abstractText | Standard genetic approaches allow the production of protein composites by fusion of polypeptides in head-to-tail fashion. Some applications would benefit from constructions that are genetically impossible, such as the site-specific linkage of proteins via their N or C termini, when a remaining free terminus is required for biological activity. We developed a method for the production of N-to-N and C-to-C dimers, with full retention of the biological activity of both fusion partners and without inflicting chemical damage on the proteins to be joined. We use sortase A to install on the N or C terminus of proteins of interest the requisite modifications to execute a strain-promoted copper-free cycloaddition and show that the ensuing ligation proceeds efficiently. Applied here to protein-protein fusions, the method reported can be extended to connecting proteins with any entity of interest. |
