Title: | Molecular quantification of Saccharomyces cerevisiae alpha-pheromone secretion |
Author(s): | Rogers DW; McConnell E; Greig D; |
Address: | "Max Planck Institute for Evolutionary Biology, Plon, Germany. rogers@evolbio.mpg.de" |
DOI: | 10.1111/j.1567-1364.2012.00817.x |
ISSN/ISBN: | 1567-1364 (Electronic) 1567-1356 (Linking) |
Abstract: | "Saccharomyces cerevisiae yeast cells court each other by producing an attractive sex pheromone specific to their mating type. Cells detect the sex pheromone from potential mates using a well-defined intracellular signalling cascade that has become a model for studying signal transduction. In contrast, the factors contributing to the production of pheromone itself are poorly characterized, despite the widespread use of the S. cerevisiae alpha-pheromone secretion pathway in industrial fungal protein expression systems. Progress in understanding pheromone secretion has been hindered by a lack of a precise and quantitative pheromone production assay. Here, we present an ELISA-based method for the quantification of alpha-pheromone secretion. In the absence of pheromone from the opposite mating type, we found that each cell secretes over 550 mature alpha-pheromone peptides per second; 90% of this total was produced from MF alpha1. The addition of a-pheromone more than doubled total alpha-pheromone secretion. This technique offers several improvements on current methods for measuring alpha-pheromone production and will allow detailed investigation of the factors regulating pheromone production in yeast" |
Keywords: | Enzyme-Linked Immunosorbent Assay/methods Mating Factor Mycology/*methods Peptides/*analysis Saccharomyces cerevisiae/*genetics/*metabolism; |
Notes: | "MedlineRogers, David W McConnell, Ellen Greig, Duncan eng Evaluation Study England 2012/06/08 FEMS Yeast Res. 2012 Sep; 12(6):668-74. doi: 10.1111/j.1567-1364.2012.00817.x. Epub 2012 Jun 25" |