Characterization of theHansenula polymorpha CPY gene encoding carboxypeptidase Y

✍ Scribed by Bellu, A. R.; van der Klei, I. J.; Rechinger, K. B.; Yavuz, M.; Veenhuis, M.; Kiel, J. A. K. W.

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 304 KB
• Volume: 15
• Category: Article
• ISSN: 0749-503X
• DOI: 10.1002/(sici)1097-0061(199902)15:3<181::aid-yea355>3.0.co;2-y

✦ Synopsis

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 cpy 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.