Applying selenocysteine-mediated expressed protein ligation to prepare the membrane enzyme selenoprotein S

Methods Enzymol. 2022:662:159-185. doi: 10.1016/bs.mie.2021.10.023. Epub 2021 Dec 24.

Abstract

The intrinsically disordered membrane-bound selenoprotein s (selenos) takes part in the protein quality control pathway, vesicle trafficking, and NF-kB signaling. The reactive selenocysteine (Sec) at the penultimate position is responsible for its enzymatic activity. We report the preparation of the soluble segment as well as the full-length selenos using expressed protein ligation. This chapter discusses the practical considerations of expressed protein ligation using selenopeptides and describes our optimized procedure for the semi-synthesis of selenos.

Keywords: Expressed protein ligation; Native chemical ligation; SEPS1; Selenocysteine; Selenopeptide; Selenoproteins; Selenos; Sels; VIMP.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Selenocysteine* / metabolism
  • Selenoproteins* / genetics
  • Selenoproteins* / metabolism

Substances

  • Selenoproteins
  • Selenocysteine