Stabilization of the CD81 Large Extracellular Loop with <i>De Novo</i> Disulfide Bonds Improves Its Amenability for Peptide Grafting.

ABSTRACT: Tetraspan proteins are significantly enriched in the membranes of exosomal vesicles (EVs) and their extracellular domains are attractive targets for engineering towards specific antigen recognition units. To enhance the tolerance of a tetraspanin fold to modification, we achieved significant thermal stabilization of the human CD81 large extracellular loop (hCD81 LEL) via de novo disulfide bonds. The best mutants were shown to exhibit a positive shift in the melting temperature (T_m) of up to 25 °C. The combination of two most potent disulfide bonds connecting different strands of the protein resulted in a mutant with a T_m of 109 °C, 43 °C over the T_m of the wild-type hCD81 LEL. A peptide sequence binding to the human transferrin r

SUBMITTER: Vogt S 

PROVIDER: S-EPMC6160918 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature