JSEMAT  Vol.3 No.4 A , October 2013
Interaction between Peptide Pheromone or Its Truncated Derivatives and Pheromone Receptor of the Fission Yeast Schizosaccharomyces pombe Examined by a Force Spectroscopy Study and a GFP Reporter Assay
ABSTRACT

In our previous study, the specific interaction between P-factor, a peptide pheromone and its receptor, Mam2, on the cell surface of the fission yeast Schizosaccharomyces pombe was investigated by two methods, an atomic force microscope (AFM) and a GFP reporter assay. The removal of Leu at C-terminal of P-factor resulted in an inactivation of P-factor function to bind Mam2 and induce the signal transduction pathway. Here, we used truncated P-factor derivatives lacking N-terminal of P-factor (P12 ~ P22: 12 ~ 22 amino acid residues from C-terminal) as ligands for Mam2. From the dose-dependent analysis of the GFP reporter assay ranging from 1 nM to 100 μM of the peptide concentration, the peptides can be classified into three groups based on EC50 and maximal GFP production level, group1 (P-factor), group2 (P17 ~ 22), and group3 (P12 ~ P16). At 0.1 μM, only P-factor induced the signal transduction pathway. At 1 μM, peptides from group2 partially induced the pathway and peptides from group3 induced the pathway a little. At 10 μM, all peptides induced the pathway mostly depending on the length of peptides. We also performed AFM experiments using P-factor and peptides from group3 to investigate the interaction between the peptides and Mam2 for comparison between the two methods.


Cite this paper
S. Hidaka, O. Nikaido, S. Kiyosaki, A. Ikai and T. Osada, "Interaction between Peptide Pheromone or Its Truncated Derivatives and Pheromone Receptor of the Fission Yeast Schizosaccharomyces pombe Examined by a Force Spectroscopy Study and a GFP Reporter Assay," Journal of Surface Engineered Materials and Advanced Technology, Vol. 3 No. 4, 2013, pp. 36-42. doi: 10.4236/jsemat.2013.34A1005.
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