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


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.

Share and Cite:

Hidaka, S. , Nikaido, O. , Kiyosaki, S. , Ikai, A. and Osada, T. (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. Journal of Surface Engineered Materials and Advanced Technology, 3, 36-42. doi: 10.4236/jsemat.2013.34A1005.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] R. Fredriksson, M. C. Lagerstrom, L.-G. Lundin and H. B. Schioth, “The G-Protein-Coupled Receptors in the Human Genome Form Five Main Families. Phylogenetic Analysis, Paralogon Groups and Fingerprints,” Molecular Pharmacology, Vol. 63, No. 6, 2003, pp. 1256-1272.
[2] S. Takeda, S. Kadowaki, T. Haga, H. Takaesu and S. Mitakud, “Identification of G Protein-Coupled Receptor Genes from the Human Genome Sequence,” FEBS Letters, Vol. 520, No. 1-3, 2002, pp. 97-101.
[3] R. Heilker, M. Wolff, C. S. Tautermann and M. Bieler, “G-Protein-Coupled Receptor-Focused Drug Discovery Using a Target Class Platform Approach,” Drug Discovery Today, Vol. 14, No. 5-6, 2009, pp. 231-240.
[4] G. Ladds, A. Goddard and J. Davey, “Functional Analysis of Heterologous GPCR Signalling Pathways in Yeast,” Trends in Biotechnology, Vol. 23, No. 7, 2005, pp. 367-373.
[5] J. Kurjan, “The Pheromone Response Pathway in Saccharomyces cerevisiae,” Annual Review of Genetics, Vol. 27, 1993, pp. 147-179.
[6] J. Davey, “Fusion of a Fission Yeast,” Yeast, Vol. 14, No. 16, 1998, pp. 1529-1566.<1529::AID-YEA357>3.0.CO;2-0
[7] S. J. Dowell and A. J. Brown, “Yeast Assays for G-Protein-Coupled Receptors,” Receptors and Channels, Vol. 552, No. 5-6, 2002, pp. 343-352.
[8] O. Nielsen, “Signal Transduction during Mating and Meiosis in S. pombe,” Trends in Cell Biology, Vol. 3, No. 2, 1993, pp. 60-65.
[9] Y. Fukui, Y. Kaziro and M. Yamamoto, “Mating PheroMone-Like Diffusible Factor Released by Schizosaccharomyces pombe,” EMBO Journal, Vol. 5, No. 8, 1986, pp. 1991-1993.
[10] K. Kitamural and C. Shimoda, “The Schizosaccharomyces Pombe Mam2 Gene Encode a Putative Pheromone Receptor Which Has a Significant Homology with the Saccharomyces cerevisiae Ste2 Protein,” The EMBO Journal, Vol. 10, No. 12, 1991, pp. 3743-3751.
[11] K. Tanaka, J. Davey, Y. Imai and M. Yamamoto, “Schizosaccharomyces pombe map3+ Encodes the Putative M-Factor Receptor,” Molecular and Cellular Biology, Vol. 13, No. 1, 1993, pp. 80-88.
[12] Y. Imai and M. Yamamoto, “The Fission Yeast Mating Pheromone P-Factor: Its Molecular Structure, Gene Structure, and Ability to Induce Gene Expression and G1 Arrest in the Mating Partner,” Genes & Development, Vol. 8, No. 3, 1994, pp. 328-338.
[13] J. Davey, “Mating Pheromones of the Fission Yeast Schizosaccharomyces pombe: Purification and Structural Characterization of M-Factor and Isolation and Analysis of Two Genes Encoding the Pheromone,” The EMBO Journal, Vol. 11, No. 3, 1992, pp. 951-960.
[14] M. Whiteway, L. Hougan, D. Dignard, D. Y. Thomas, L. Bell, G. C. Saari, F. J. Grant, P. O’Hara and V. L. MacKay, “The STE4 and STE18 Genes of Yeast Encode Potential β and γ Subunits of the Mating Factor Receptor-Coupled G Protein,” Cell, Vol. 56, No. 3, 1989, pp. 467-477.
[15] T. Obara, M. Nakafuku, M. Yamamoto and Y. Kaziro, “Isolation and Characterization of a Gene Encoding a G-Protein a Subunit from Schizosaccharomyces pombe: Involvement in Mating and Sporulation Pathways,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 88, No. 13, 1991, pp. 5877-5881.
[16] E. J. Neer “Heterotrimeric G Proteins: Organizers of Transmembrane Signals,” Cell, Vol. 80, No. 2, 1995, pp. 249-257.
[17] H. E. Hamm, “The Many Faces of G Protein Signaling,” The Journal of Biological Chemistry, Vol. 273, No. 2, 1998, pp. 669-672.
[18] M. Yamamoto, “Regulation of Meiosis in Fission Yeast,” Cell Structure and Function, Vol. 21, No. 5, 1996, pp. 431-436.
[19] G. Binnig and H. Rohrer, “Scanning Tunneling Microscopy,” Surface Science, Vol. 126, No. 1-3, 1983, pp. 236-244.
[20] M. Radmacher, M. Fritz, J. P. Cleveland, D. A. Walters and P. K. Hansma, “Imaging Adhesion Forces and Elasticity of Lysozyme Adsorbed on Mica with the Atomic Force Microscope,” Langmuir, Vol. 10, No. 10, 1994, pp. 3809-3814.
[21] K. Mitsui, M. Hara and A. Ikai, “Mechanical Unfolding of a2-Macroglobulin Molecules with Atomic Force Microscope,” FEBS Letters, Vol. 385, No. 1-2, 1996, pp. 29-33.
[22] M. Rief, M. Gautel, F. Oesterhelt, J. M. Fernandez and H. E. Gaub, “Reversible Unfolding of Individual Titin Immunoglobulin Domains by AFM,” Science, Vol. 276, No. 5315, 1997, pp. 1109-1112.
[23] J. P. Michel, I. L. Ivanovska, M. M. Gibbons, et al., “Nanoindentation Studies of Full and Empty Viral Capsids and the Effects of Capsid Protein Mutations on Elasticity and Strength,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 103, No. 16, 2006, pp. 6184-6189.
[24] G. Lee, K. Abdi, Y. Jiang, P. Michaely, V. Bennett and P. E. Marszalek, “Nanospring Behaviour of Ankyrin Repeats,” Nature, Vol. 440, No. 7081, 2006, pp. 246-249.
[25] R. Afrin, T. Yamada and A. Ikai, “Analysis of Force Curves Obtained on the Live Cell Membrane Using Chemically Modified AFM Probes,” Ultramicroscopy, Vol. 100, No. 3-4, 2004, pp. 187-195.
[26] C. Lesoil, T. Nonaka, H. Sekiguchi, et al., “Molecular Shape and Binding Force of Mycoplasma Mobile’s Leg Protein Gli349 Revealed by an AFM Study,” Biochemical and Biophysical Research Communications, Vol. 391, No. 3, 2010, pp. 1312-1317.
[27] A. Yersin, T. Osada and A. Ikai, “Exploring Transferrin-Receptor Interactions at the Single-Molecule Level,” Biophysical Journal, Vol. 94, No. 1, 2008, pp. 230-240.
[28] S. Sasuga, R. Abe, O. Nikaido, S. Kiyosaki, H. Sekiguchi, A. Ikai and T. Osada, “Interaction between Pheromone and Its Receptor of the Fission Yeast Schizosaccharomyces pombe Examined by a Force Spectroscopy Study,” Journal of Biomedicine and Biotechnology, Vol. 2012, 2012, Article ID: 804793.
[29] H. Kim, F. Asgari, M. Kato-Negishi, S. Ohkura, H. Okamura, H. Arakawa, T. Osada and A. Ikai, “Distribution of Olfactory Marker Protein on a Tissue Section of Vomeronasal Organ Measured by AFM,” Colloids and Surfaces B: Biointerfaces, Vol. 61, No. 2, 2008, pp. 311-314.
[30] H. Kim, H. Arakawa, N. Hatae, Y. Sugimoto, O. Matsumoto, T. Osada, A. Ichikawa and A. Ikai, “Quantification of the Number of EP3 Receptors on a Living CHO Cell Surface by the AFM,” Ultramicroscopy, Vol. 106, No. 8-9, 2006, pp. 652-662.

Copyright © 2022 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.