Characterization of the Hansenula polymorpha CPY gene encoding carboxypeptidase Y
We have isolated the Hansenula polymorpha CPY gene encoding carboxypeptidase Y (Hp‐CPY). The deduced amino acid sequence revealed that Hp‐CPY consists of 541 amino acids and has a calculated Mr of 60,793. The protein is highly similar to Saccharomyces cerevisiae CPY (61·8% identity). At the N‐termin...
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| Veröffentlicht in: | Yeast (Chichester, England) England), 1999-02, Vol.15 (3), p.181-189 |
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| creator | Bellu, A. R. van der Klei, I. J. Rechinger, K. B. Yavuz, M. Veenhuis, M. Kiel, J. A. K. W. |
| description | We have isolated the Hansenula polymorpha CPY gene encoding carboxypeptidase Y (Hp‐CPY). The deduced amino acid sequence revealed that Hp‐CPY consists of 541 amino acids and has a calculated Mr of 60,793. The protein is highly similar to Saccharomyces cerevisiae CPY (61·8% identity). At the N‐terminus of Hp‐CPY signals for the entry into the secretory pathway and subsequent sorting to the vacuole were identified. Immunocytochemically, using monospecific antibodies raised against Hp‐CPY, the protein was localized to the vacuole. On Western blots, a diffuse protein band was observed in extracts of H. polymorpha cells, suggesting that the protein is glycosylated. This was confirmed by endoglycosidase H treatment, which resulted in a strong reduction of the apparent Mr of the protein. We have investigated the effect of CPY deletion on the degradation of peroxisomes, an autophagous process that occurs when the organelles become redundant for growth. In Δcpy cells peroxisomal proteins were degraded in the vacuole as efficiently as in wild‐type H. polymorpha cells, indicating that CPY is not a major proteinase in this pathway. Copyright © 1999 John Wiley & Sons, Ltd. |
| doi_str_mv | 10.1002/(SICI)1097-0061(199902)15:3<181::AID-YEA355>3.0.CO;2-Y |
| format | Article |
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R. ; van der Klei, I. J. ; Rechinger, K. B. ; Yavuz, M. ; Veenhuis, M. ; Kiel, J. A. K. W.</creator><creatorcontrib>Bellu, A. R. ; van der Klei, I. J. ; Rechinger, K. B. ; Yavuz, M. ; Veenhuis, M. ; Kiel, J. A. K. W.</creatorcontrib><description>We have isolated the Hansenula polymorpha CPY gene encoding carboxypeptidase Y (Hp‐CPY). The deduced amino acid sequence revealed that Hp‐CPY consists of 541 amino acids and has a calculated Mr of 60,793. The protein is highly similar to Saccharomyces cerevisiae CPY (61·8% identity). At the N‐terminus of Hp‐CPY signals for the entry into the secretory pathway and subsequent sorting to the vacuole were identified. Immunocytochemically, using monospecific antibodies raised against Hp‐CPY, the protein was localized to the vacuole. On Western blots, a diffuse protein band was observed in extracts of H. polymorpha cells, suggesting that the protein is glycosylated. This was confirmed by endoglycosidase H treatment, which resulted in a strong reduction of the apparent Mr of the protein. We have investigated the effect of CPY deletion on the degradation of peroxisomes, an autophagous process that occurs when the organelles become redundant for growth. In Δcpy cells peroxisomal proteins were degraded in the vacuole as efficiently as in wild‐type H. polymorpha cells, indicating that CPY is not a major proteinase in this pathway. 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R.</creatorcontrib><creatorcontrib>van der Klei, I. J.</creatorcontrib><creatorcontrib>Rechinger, K. B.</creatorcontrib><creatorcontrib>Yavuz, M.</creatorcontrib><creatorcontrib>Veenhuis, M.</creatorcontrib><creatorcontrib>Kiel, J. A. K. W.</creatorcontrib><title>Characterization of the Hansenula polymorpha CPY gene encoding carboxypeptidase Y</title><title>Yeast (Chichester, England)</title><addtitle>Yeast</addtitle><description>We have isolated the Hansenula polymorpha CPY gene encoding carboxypeptidase Y (Hp‐CPY). The deduced amino acid sequence revealed that Hp‐CPY consists of 541 amino acids and has a calculated Mr of 60,793. The protein is highly similar to Saccharomyces cerevisiae CPY (61·8% identity). At the N‐terminus of Hp‐CPY signals for the entry into the secretory pathway and subsequent sorting to the vacuole were identified. Immunocytochemically, using monospecific antibodies raised against Hp‐CPY, the protein was localized to the vacuole. On Western blots, a diffuse protein band was observed in extracts of H. polymorpha cells, suggesting that the protein is glycosylated. This was confirmed by endoglycosidase H treatment, which resulted in a strong reduction of the apparent Mr of the protein. We have investigated the effect of CPY deletion on the degradation of peroxisomes, an autophagous process that occurs when the organelles become redundant for growth. In Δcpy cells peroxisomal proteins were degraded in the vacuole as efficiently as in wild‐type H. polymorpha cells, indicating that CPY is not a major proteinase in this pathway. Copyright © 1999 John Wiley & Sons, Ltd.</description><subject>Amino Acid Sequence</subject><subject>autophagy</subject><subject>Blotting, Western</subject><subject>Carboxypeptidases - chemistry</subject><subject>Carboxypeptidases - genetics</subject><subject>Carboxypeptidases - metabolism</subject><subject>Cathepsin A</subject><subject>Cloning, Molecular</subject><subject>Genes, Fungal - genetics</subject><subject>Genomic Library</subject><subject>Glycoside Hydrolases - metabolism</subject><subject>Glycosylation</subject><subject>Hansenula polymorpha</subject><subject>Immunohistochemistry</subject><subject>methylotrophic yeast</subject><subject>Microbodies - metabolism</subject><subject>Microbodies - physiology</subject><subject>Molecular Sequence Data</subject><subject>Molecular Weight</subject><subject>Mutagenesis, Insertional</subject><subject>Open Reading Frames - genetics</subject><subject>peroxisome turnover</subject><subject>Pichia - cytology</subject><subject>Pichia - enzymology</subject><subject>Pichia - genetics</subject><subject>Pichia - growth & development</subject><subject>Protein Sorting Signals - genetics</subject><subject>Recombinant Fusion Proteins - chemistry</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Sequence Alignment</subject><subject>Sequence Deletion</subject><subject>Sequence Homology, Nucleic Acid</subject><subject>vacuolar proteinases</subject><subject>Vacuoles - metabolism</subject><issn>0749-503X</issn><issn>1097-0061</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1999</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpNkdtKw0AQhhdRtFZfQfZK6kXqzm43m1QplHgqFKqoYK6W7WbSRtIk5oDWpzelVbwamPlmBv6PkBGwPjDGL3vPk2ByAcxXDmMu9MD3fcYvQA7FNXgwHI4nN054OxZSjkSf9YPZFXfCPdL5W9knHaYGviOZeDsix1X1zhiA5N4hOWpfKAWe7JCnYGlKY2ssk29TJ3lG85jWS6QPJqswa1JDizxdr_KyWBoaPIZ0gRlSzGweJdmCWlPO8691gUWdRKZCGp6Qg9ikFZ7uape83t2-BA_OdHY_CcZTp-BiIB3jcaliIS0Y6wrXdTEegGon88j61rrxYC5iwFh4PkbG9VUkmIfWGCFQKQuiS863d4sy_2iwqvUqqSymqckwbyoNijOQntuCZzuwma8w0kWZrEy51r8htMDbFvhMUlz_m28Qrjcm9CZWvYlVb01okFro1oRuReitiLbBdDDTXIe7jvgBsyOBog</recordid><startdate>199902</startdate><enddate>199902</enddate><creator>Bellu, A. R.</creator><creator>van der Klei, I. J.</creator><creator>Rechinger, K. B.</creator><creator>Yavuz, M.</creator><creator>Veenhuis, M.</creator><creator>Kiel, J. A. K. W.</creator><general>John Wiley & Sons, Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>8FD</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>199902</creationdate><title>Characterization of the Hansenula polymorpha CPY gene encoding carboxypeptidase Y</title><author>Bellu, A. R. ; van der Klei, I. J. ; Rechinger, K. B. ; Yavuz, M. ; Veenhuis, M. ; Kiel, J. A. K. W.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p2345-a8257f35c1ac63666ef417345bdc9cc6f4b3f1ef389eda697d308ecaa33e77c13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1999</creationdate><topic>Amino Acid Sequence</topic><topic>autophagy</topic><topic>Blotting, Western</topic><topic>Carboxypeptidases - chemistry</topic><topic>Carboxypeptidases - genetics</topic><topic>Carboxypeptidases - metabolism</topic><topic>Cathepsin A</topic><topic>Cloning, Molecular</topic><topic>Genes, Fungal - genetics</topic><topic>Genomic Library</topic><topic>Glycoside Hydrolases - metabolism</topic><topic>Glycosylation</topic><topic>Hansenula polymorpha</topic><topic>Immunohistochemistry</topic><topic>methylotrophic yeast</topic><topic>Microbodies - metabolism</topic><topic>Microbodies - physiology</topic><topic>Molecular Sequence Data</topic><topic>Molecular Weight</topic><topic>Mutagenesis, Insertional</topic><topic>Open Reading Frames - genetics</topic><topic>peroxisome turnover</topic><topic>Pichia - cytology</topic><topic>Pichia - enzymology</topic><topic>Pichia - genetics</topic><topic>Pichia - growth & development</topic><topic>Protein Sorting Signals - genetics</topic><topic>Recombinant Fusion Proteins - chemistry</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Sequence Alignment</topic><topic>Sequence Deletion</topic><topic>Sequence Homology, Nucleic Acid</topic><topic>vacuolar proteinases</topic><topic>Vacuoles - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bellu, A. R.</creatorcontrib><creatorcontrib>van der Klei, I. J.</creatorcontrib><creatorcontrib>Rechinger, K. B.</creatorcontrib><creatorcontrib>Yavuz, M.</creatorcontrib><creatorcontrib>Veenhuis, M.</creatorcontrib><creatorcontrib>Kiel, J. A. K. W.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Yeast (Chichester, England)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bellu, A. R.</au><au>van der Klei, I. J.</au><au>Rechinger, K. B.</au><au>Yavuz, M.</au><au>Veenhuis, M.</au><au>Kiel, J. A. K. W.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of the Hansenula polymorpha CPY gene encoding carboxypeptidase Y</atitle><jtitle>Yeast (Chichester, England)</jtitle><addtitle>Yeast</addtitle><date>1999-02</date><risdate>1999</risdate><volume>15</volume><issue>3</issue><spage>181</spage><epage>189</epage><pages>181-189</pages><issn>0749-503X</issn><eissn>1097-0061</eissn><abstract>We have isolated the Hansenula polymorpha CPY gene encoding carboxypeptidase Y (Hp‐CPY). The deduced amino acid sequence revealed that Hp‐CPY consists of 541 amino acids and has a calculated Mr of 60,793. The protein is highly similar to Saccharomyces cerevisiae CPY (61·8% identity). At the N‐terminus of Hp‐CPY signals for the entry into the secretory pathway and subsequent sorting to the vacuole were identified. Immunocytochemically, using monospecific antibodies raised against Hp‐CPY, the protein was localized to the vacuole. On Western blots, a diffuse protein band was observed in extracts of H. polymorpha cells, suggesting that the protein is glycosylated. This was confirmed by endoglycosidase H treatment, which resulted in a strong reduction of the apparent Mr of the protein. We have investigated the effect of CPY deletion on the degradation of peroxisomes, an autophagous process that occurs when the organelles become redundant for growth. In Δcpy cells peroxisomal proteins were degraded in the vacuole as efficiently as in wild‐type H. polymorpha cells, indicating that CPY is not a major proteinase in this pathway. Copyright © 1999 John Wiley & Sons, Ltd.</abstract><cop>Chichester, UK</cop><pub>John Wiley & Sons, Ltd</pub><pmid>10077185</pmid><doi>10.1002/(SICI)1097-0061(199902)15:3<181::AID-YEA355>3.0.CO;2-Y</doi><tpages>9</tpages></addata></record> |
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| subjects | Amino Acid Sequence autophagy Blotting, Western Carboxypeptidases - chemistry Carboxypeptidases - genetics Carboxypeptidases - metabolism Cathepsin A Cloning, Molecular Genes, Fungal - genetics Genomic Library Glycoside Hydrolases - metabolism Glycosylation Hansenula polymorpha Immunohistochemistry methylotrophic yeast Microbodies - metabolism Microbodies - physiology Molecular Sequence Data Molecular Weight Mutagenesis, Insertional Open Reading Frames - genetics peroxisome turnover Pichia - cytology Pichia - enzymology Pichia - genetics Pichia - growth & development Protein Sorting Signals - genetics Recombinant Fusion Proteins - chemistry Recombinant Fusion Proteins - metabolism Sequence Alignment Sequence Deletion Sequence Homology, Nucleic Acid vacuolar proteinases Vacuoles - metabolism |
| title | Characterization of the Hansenula polymorpha CPY gene encoding carboxypeptidase Y |
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