TITLE:
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
AUTHORS:
Sho Hidaka, Osamu Nikaido, Shoichi Kiyosaki, Atsushi Ikai, Toshiya Osada
KEYWORDS:
GPCR; Mam2; Pheromone; GFP; AFM; Yeast
JOURNAL NAME:
Journal of Surface Engineered Materials and Advanced Technology,
Vol.3 No.4A,
October
22,
2013
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.