Toward the semisynthesis of multidomain transmembrane receptors: Modification of Eph tyrosine kinases Journal Article
| Authors: | Singla, N.; Himanen, J. P.; Muir, T. W.; Nikolov, D. B. |
| Article Title: | Toward the semisynthesis of multidomain transmembrane receptors: Modification of Eph tyrosine kinases |
| Abstract: | Expressed protein ligation (EPL) is a protein engineering approach that allows the modification or assembly of a target protein from multiple recombinant and synthetic polypeptides. EPL has been previously used to modify intracellular proteins and small integral membrane proteins for structural and functional studies. Here we describe the semisynthetic site-specific modification of the complete, multidomain extracellular regions of both A and B classes of Eph receptor tyrosine kinases. We show that the ectodomains of these receptors can be ligated to different peptides under carefully established experimental conditions, while their biological activity is retained. This work extends the boundaries of the EPL technique for semisynthesis of multidomain, extracellular, disulfide-bonded, and glycosylated proteins and highlights its potential application for reconstituting entire single-pass transmembrane proteins. Copyright © 2008 The Protein Society. |
| Keywords: | protein expression; human cell; nonhuman; protein domain; protein function; animal cell; animals; cell line; protein tyrosine kinase; cloning, molecular; recombinant fusion proteins; protein synthesis; peptide fragments; recombinant protein; peptides; protein structure, tertiary; protein structure; receptors, eph family; protein modification; polypeptide; expressed protein ligation; ephrins; ephrin receptor; membrane receptor; eph receptor tyrosine kinase; inteins; multidomain extracellular region; glycosylated protein; protein engineering |
| Journal Title: | Protein Science |
| Volume: | 17 |
| Issue: | 10 |
| ISSN: | 0961-8368 |
| Publisher: | Wiley Blackwell |
| Date Published: | 2008-10-01 |
| Start Page: | 1740 |
| End Page: | 1747 |
| Language: | English |
| DOI: | 10.1110/ps.035659.108 |
| PUBMED: | 18628240 |
| PROVIDER: | scopus |
| PMCID: | PMC2548361 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 3" - "Export Date: 17 November 2011" - "CODEN: PRCIE" - "Source: Scopus" |
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