Identification and characterization of polycystin-2, the PKD2 gene product

Yiqiang Cai, Yoshiko Maeda, Anna Cedzich, Vicente E. Torres, Guanqing Wu, Tomohito Hayashi, Toshio Mochizuki, Jong Hoon Park, Ralph Witzgall, Stefan Somlo

Research output: Contribution to journalArticle

276 Scopus citations

Abstract

PKD2, the second gene for the autosomal dominant polycystic kidney disease (ADPKD), encodes a protein, polycystin-2, with predicted structural similarity to cation channel subunits. However, the function of polycystin-2 remains unknown. We used polyclonal antisera specific for the intracellular NH2 and COOH termini to identify polycystin-2 as an ~110-kDa integral membrane glycoprotein. Polycystin-2 from both native tissues and cells in culture is sensitive to Endo H suggesting the continued presence of high- mannose oligosaccharides typical of pre-middle Golgi proteins. Immunofluorescent cell staining of polycystin-2 shows a pattern consistent with localization in the endoplasmic reticulum. This finding is confirmed by co-localization with protein-disulfide isomerase as determined by double indirect immunofluorescence and co-distribution with calnexin in subcellular fractionation studies. Polycystin-2 translation products truncated at or after Gly821 retain their exclusive endoplasmic reticulum localization while products truncated at or before Glu787 additionally traffic to the plasma membrane. Truncation mutants that traffic to the plasma membrane acquire Endo H resistance and can be biotinylated on the cell surface in intact cells. The 34-amino acid region Glu787-Ser820, containing two putative phosphorylation sites, is responsible for the exclusive endoplasmic reticulum localization of polycystin-2 and is the site of specific interaction with an as yet unidentified protein binding partner for polycystin-2. The localization of full-length polycystin-2 to intracellular membranes raises the possibility that the PKD2 gene product is a subunit of intracellular channel complexes.

Original languageEnglish (US)
Pages (from-to)28557-28565
Number of pages9
JournalJournal of Biological Chemistry
Volume274
Issue number40
DOIs
StatePublished - Oct 1 1999

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Identification and characterization of polycystin-2, the PKD2 gene product'. Together they form a unique fingerprint.

  • Cite this

    Cai, Y., Maeda, Y., Cedzich, A., Torres, V. E., Wu, G., Hayashi, T., Mochizuki, T., Park, J. H., Witzgall, R., & Somlo, S. (1999). Identification and characterization of polycystin-2, the PKD2 gene product. Journal of Biological Chemistry, 274(40), 28557-28565. https://doi.org/10.1074/jbc.274.40.28557