Cloning and expression of a heme binding protein from the genome of Saccharomyces cerevisiae.

Karine Auclair, Hong-Wei Huang, Pierre Moënne-Loccoz, Paul R Ortiz de Montellano

Department of Chemistry, McGill University, 801 Sherbrooke Street West, Montréal, Québec, Canada H3A 2K6. karine.auclair@mcgill.ca

Protein expression and purification 2003 Apr

filter terms:

The YLR205c gene of Saccharomyces cerevisiae does not show significant sequence identity to any known gene, except for heme oxygenase (22% to human HO-1). The YLR205 ORF was cloned and overexpressed in both Escherichia coli and S. cerevisiae. Both expression systems yielded proteins that bound heme tightly. The isolated YLR205c protein underwent reduction in the presence of either NADPH-cytochrome P450 reductase or NADH-putidaredoxin-putidaredoxin reductase but did not exhibit heme oxygenase activity. The protein exhibited modest H(2)O(2)-dependent peroxidase activities with guaiacol, potassium iodide, and 2,2(')-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS). Thus, YLR205c codes for a hemoprotein of unknown physiological function that exhibits peroxidase activity.

Citation

Karine Auclair, Hong-Wei Huang, Pierre Moënne-Loccoz, Paul R Ortiz de Montellano. Cloning and expression of a heme binding protein from the genome of Saccharomyces cerevisiae. Protein expression and purification. 2003 Apr;28(2):340-9