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Updated: Jul 18, 2025
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EPR Monitored Redox Titration of the Cofactors of Saccharomyces cerevisiae Nar1
Published on: November 26, 2014
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Saccharomyces cerevisiae NRE1 and IRC24 Encode Paralogous Benzil Oxidoreductases
Brandon Garcia1, Kasandra J Riley2
1Rollins College, Winter Park, Florida, United States.
Micropublication Biology
|August 21, 2023
Summary
Researchers identified Nre1p, a paralog of Irc24p in Saccharomyces cerevisiae. Both proteins are benzil oxidoreductases, with Nre1p
Area of Science:
- Biochemistry
- Molecular Biology
- Yeast Genetics
Background:
- Irc24p is a known benzil oxidoreductase in Saccharomyces cerevisiae.
- A putative paralog, Nre1p, was identified downstream of Irc24p on chromosome IX.
- Both proteins share significant sequence identity and structural features, suggesting related functions.
Purpose of the Study:
- To characterize the biochemical function of the newly identified Nre1p.
- To compare the enzymatic activity of Nre1p with its paralog, Irc24p.
- To confirm the cofactor preference for these benzil oxidoreductases.
Main Methods:
- Bioinformatic analysis (PANTHER, PFAM) of Irc24p and Nre1p amino acid sequences.
- Structural alignment to predict conserved active sites and Rossmann folds.
- Purification of hexahistidine-tagged Irc24p and Nre1p.
- In vitro enzymatic assays to measure benzil reduction kinetics.
Main Results:
- Both Irc24p and Nre1p were purified and confirmed as benzil oxidoreductases.
- Similar catalytic kinetics were observed for both proteins in reducing benzil.
- A preference for NADPH as a cofactor was demonstrated for both enzymes.
Conclusions:
- Nre1p is functionally characterized as a benzil oxidoreductase, demonstrating in vitro activity for the first time.
- Irc24p and Nre1p represent a pair of related benzil oxidoreductases in Saccharomyces cerevisiae with similar biochemical properties.
- The findings provide insights into the functional redundancy and evolution of oxidoreductases in yeast.
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