The semisynthesis of nucleolar human selenoprotein H

Rebecca Notis Dardashti; Shay Laps; Jacob S Gichtin; Norman Metanis

doi:10.1039/d3sc03059h

The semisynthesis of nucleolar human selenoprotein H

Chem Sci. 2023 Oct 24;14(44):12723-12729. doi: 10.1039/d3sc03059h. eCollection 2023 Nov 15.

Authors

Rebecca Notis Dardashti¹, Shay Laps¹, Jacob S Gichtin¹, Norman Metanis^{1

2

3}

Affiliations

¹ The Institute of Chemistry, The Hebrew University of Jerusalem Edmond J. Safra Campus Givat Ram Jerusalem 9190401 Israel.
² Casali Center for Applied Chemistry, The Hebrew University of Jerusalem Edmond J. Safra Campus Givat Ram Jerusalem 9190401 Israel.
³ The Center for Nanoscience and Nanotechnology, The Hebrew University of Jerusalem, Edmond J. Safra Campus Givat Ram Jerusalem 9190401 Israel metanis@mail.huji.ac.il.

Abstract

The human selenoprotein H is the only selenocysteine-containing protein that is located in the cell's nucleolus. In vivo studies have suggested that it plays some role in DNA binding, consumption of reactive oxygen species, and may serve as a safeguard against cancers. However, the protein has never been isolated and, as a result, not yet fully characterized. Here, we used a semi-synthetic approach to obtain the full selenoprotein H with a S43T mutation. Using biolayer interferometry, we also show that the Cys-containing mutant of selenoprotein H is capable of binding DNA with sub-micromolar affinity. Employing state-of-the-art expressed protein ligation (EPL), our devised semi-synthetic approach can be utilized for the production of numerous, hard-to-obtain proteins of biological and therapeutic relevance.