Characterization of the biochemical and biophysical properties of the Saccharomyces cerevisiae phosphate transporter Pho89

Palanivelu Sengottaiyan, Jitka Petrlova, Jens O. Lagerstedt, Lorena Ruiz-Pavon, Madhu S. Budamagunta, John C Voss, Bengt L. Persson

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

In Saccharomyces cerevisiae, Pho89 mediates a cation-dependent transport of Pi across the plasma membrane. This integral membrane protein belongs to the Inorganic Phosphate Transporter (PiT) family, a group that includes the mammalian Na+/Pi cotransporters Pit1 and Pit2. Here we report that the Pichia pastoris expressed recombinant Pho89 was purified in the presence of Foscholine-12 and functionally reconstituted into proteoliposomes with a similar substrate specificity as observed in an intact cell system. The alpha-helical content of the Pho89 protein was estimated to 44%. EPR analysis showed that purified Pho89 protein undergoes conformational change upon addition of substrate.

Original languageEnglish (US)
Pages (from-to)551-556
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume436
Issue number3
DOIs
StatePublished - Jul 5 2013

Keywords

  • Circular dichroism
  • Oligomer
  • Pho89
  • Phosphate transport
  • Pichia pastoris
  • Reconstitution

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

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