Ergothioneine in an Enzyme: Using Protein Engineering to Create Unique Antioxidant Enzymes Containing 2-thiohistidine
Conference Year
2023
Abstract
Ergothioneine is a natural sulfur containing amino acid and a unique antioxidant compound. Ergothioneine is unable to be incorporated into a peptide because its nitrogen is trimethylated; however, 2-thiohistidine is an analogue of ergothioneine with similar antioxidant properties that can be inserted into a peptide/protein. This project utilizes protein engineering to replace a catalytic cysteine residue of Plasmodium falciparum thioredoxin reductase (PfTrxR)with 2-thiohistidine and investigates gains in function in three main mutants: PfTrxR-CGGGK2THG, PfTrxR-CUGGK2THG, and PFTrxR-CG2THKPG2THK. Our data supports the protective effects of 2-thiohistidine on the redox-active residue selenocysteine and the potential conferral of the ability to metabolize 1O2.
Primary Faculty Mentor Name
Robert Hondal
Status
Undergraduate
Student College
College of Arts and Sciences
Second Student College
Patrick Leahy Honors College
Program/Major
Biochemistry
Primary Research Category
Physical Science
Ergothioneine in an Enzyme: Using Protein Engineering to Create Unique Antioxidant Enzymes Containing 2-thiohistidine
Ergothioneine is a natural sulfur containing amino acid and a unique antioxidant compound. Ergothioneine is unable to be incorporated into a peptide because its nitrogen is trimethylated; however, 2-thiohistidine is an analogue of ergothioneine with similar antioxidant properties that can be inserted into a peptide/protein. This project utilizes protein engineering to replace a catalytic cysteine residue of Plasmodium falciparum thioredoxin reductase (PfTrxR)with 2-thiohistidine and investigates gains in function in three main mutants: PfTrxR-CGGGK2THG, PfTrxR-CUGGK2THG, and PFTrxR-CG2THKPG2THK. Our data supports the protective effects of 2-thiohistidine on the redox-active residue selenocysteine and the potential conferral of the ability to metabolize 1O2.