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Processing and maturation of carboxypeptidase Y and alkaline phosphatase in Schizosaccharomyces pombe

  • Applied Genetics and Molecular Biotechnology
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Abstract

Schizosaccharomyces pombe carboxypeptidase Y (CPY) is synthesized as a zymogen and transported into the vacuole where maturation and activation occurs. The 110-kDa S. pombe CPY precursor is processed twice and finally converted to a mature form consisting of polypeptides of approximately 19 and 32 kDa linked by a single disulfide bond. In Saccharomyces cerevisiae, maturation of CPY occurs mostly through the activity of vacuolar aspartyl protease Pep4p, whereas a Pep4p homolog has not been found in the S. pombe genome database. Based on analysis of protease-deficient mutants, we found that S. pombe CPY was not able to be processed or activated in isp6Δpsp3Δ double disruptants. Both Isp6p and Psp3p are subtilase-type serine proteases with related sequences. Moreover, alkaline phosphatase of S. pombe was found to be localized at the vacuolar membrane and was also unprocessed in isp6Δpsp3Δ double disruptants. Vacuolar localization of GFP-fused Isp6p and Psp3p was determined by fluorescence microscopy. These results suggest that the two serine proteases Isp6p and Psp3p are functional in the vacuole and are involved in proteolytic processing of vacuolar proteins.

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Acknowledgments

We thank Yoko Kusunoki, Yukio Ozaka, and Naotaka Tanaka for excellent technical assistance. This work was partly supported by the Project for Development of a Technological Infrastructure for Industrial Bioprocesses on R&D of New Industrial Science and Technology Frontiers by the Ministry of Economy, Trade & Industry, as supported by the New Energy and Industrial Technology Development Organization.

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Authors and Affiliations

  1. ASPEX Division, Research Center, Asahi Glass Co., Ltd., 1150 Hazawa-cho, Kanagawa, 221-8755, Japan

    Hiroyuki Mukaiyama, Tomoko Iwaki & Alimjan Idiris

  2. Department of Bioscience & Biotechnology, Faculty of Agriculture, Kyushu University, Hakozaki 6-10-1, Fukuoka, 812-8581, Japan

    Hiroyuki Mukaiyama & Kaoru Takegawa

  3. Department of Life Sciences, Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa, 761-0795, Japan

    Hiroyuki Mukaiyama, Tomoko Iwaki & Kaoru Takegawa

  4. Laboratory of Applied Microbiology, Department of Bioscience & Biotechnology, Faculty of Agriculture, Kyushu University, Hakozaki 6-10-1, Fukuoka, 812-8581, Japan

    Kaoru Takegawa

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  1. Hiroyuki Mukaiyama
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  2. Tomoko Iwaki
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Correspondence to Kaoru Takegawa.

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Mukaiyama, H., Iwaki, T., Idiris, A. et al. Processing and maturation of carboxypeptidase Y and alkaline phosphatase in Schizosaccharomyces pombe . Appl Microbiol Biotechnol 90, 203–213 (2011). https://doi.org/10.1007/s00253-010-3031-3

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  • DOI: https://doi.org/10.1007/s00253-010-3031-3

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