Calcium-assisted sortase A cleavage of SUMOylated metallothionein constructs leads to high-yield production of human MT3.

ABSTRACT:

Background

Mammalian metallothioneins (MTs) are small (6-7 kDa), intracellular, cysteine-rich, metal-binding proteins involved, inter alia, in the homeostasis of zinc and copper, detoxification of heavy metals, antioxidation against reactive oxygen species, and protection against DNA damage. The high cysteine content (~ 30%) in MTs makes them toxic to bacterial cells during protein production, resulting in low yield. To address this issue, we present for the first time a combinatorial approach using the small ubiquitin-like modifier (SUMO) and/or sortase as fusion tags for high-level expression of human MT3 in E. coli and its purification by three different strategies.

Results

Three different plasmids were generated using SUMO, sortase A pentamutant (eSrtA), and sortase reco

SUBMITTER: Singh AK 

PROVIDER: S-EPMC10334657 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature