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Toward the semisynthesis of multidomain transmembrane receptors: Modification of Eph tyrosine kinases
Author(s) -
Singla Nikhil,
Himanen Juha Pekka,
Muir Tom W.,
Nikolov Dimitar B.
Publication year - 2008
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.035659.108
Subject(s) - semisynthesis , biochemistry , transmembrane protein , receptor tyrosine kinase , erythropoietin producing hepatocellular (eph) receptor , microbiology and biotechnology , chemistry , transmembrane domain , g protein coupled receptor , integral membrane protein , receptor protein tyrosine kinases , receptor , membrane protein , biology , membrane
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.