title: Characterization of the biochemical and biophysical properties of the Saccharomyces cerevisiae phosphate transporter Pho89.
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.
authors.affiliation: Linnaeus University Centre for Biomaterials Chemistry, Department of Chemistry and Biomedical Sciences, Linnaeus University, SE-391 82 Kalmar, Sweden.
authors.forename: Palanivelu
authors.initials: P
authors.lastname: Sengottaiyan
authors.forename: Jitka
authors.initials: J
authors.lastname: Petrlova
authors.forename: Jens O
authors.initials: JO
authors.lastname: Lagerstedt
authors.forename: Lorena
authors.initials: L
authors.lastname: Ruiz-Pavon
authors.forename: Madhu S
authors.initials: MS
authors.lastname: Budamagunta
authors.forename: John C
authors.initials: JC
authors.lastname: Voss
authors.forename: Bengt L
authors.initials: BL
authors.lastname: Persson
mesh_terms: D001692:Biological Transport; D002462:Cell Membrane / Q000737:chemistry; D002942:Circular Dichroism; D004578:Electron Spin Resonance Spectroscopy; D010767:Phosphorylcholine / Q000031:analogs & derivatives / Q000737:chemistry; D010843:Pichia / Q000737:chemistry; D011485:Protein Binding; D017433:Protein Structure, Secondary; D011510:Proteolipids / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D050611:Sodium-Phosphate Cotransporter Proteins, Type III / Q000737:chemistry*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: C493567:PHO89 protein, S cerevisiae; D011510:Proteolipids; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D050611:Sodium-Phosphate Cotransporter Proteins, Type III; C020485:proteoliposomes; D010767:Phosphorylcholine; C028810:dodecylphosphocholine
keywords: CD; Circular dichroism; EPR; Oligomer; Pho89; Phosphate transport; Pichia pastoris; Reconstitution; circular dichroism; electrochemical Na(+) concentration gradient; electron paramagnetic resonance; ΔpNa(+)