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Structural modeling of dual-affinity purified Pho84 phosphate transporter

Lagerstedt, Jens O LU ; Voss, John C LU ; Wieslander, Ake and Persson, Bengt L (2004) In FEBS Letters 578(3). p.8-262
Abstract

The phosphate transporter Pho84 of Saccharomyces cerevisiae is predicted to contain 12 transmembrane (TM) regions, divided into two partially duplicated parts of 6 TM segments. The three-dimensional (3D) organization of the Pho84 protein has not yet been determined. However, the 3D crystal structure of the Escherichia coli MFS glycerol-3-phosphate/phosphate antiporter, GlpT, and lactose transporter, LacY, has recently been determined. On the basis of extensive prediction and fold recognition analyses (at the MetaServer), GlpT was proposed as the best structural template on which the arrangement of TM segments of the Pho84 transporter was fit, using the comparative structural modeling program MODELLER. To initiate an evaluation of the... (More)

The phosphate transporter Pho84 of Saccharomyces cerevisiae is predicted to contain 12 transmembrane (TM) regions, divided into two partially duplicated parts of 6 TM segments. The three-dimensional (3D) organization of the Pho84 protein has not yet been determined. However, the 3D crystal structure of the Escherichia coli MFS glycerol-3-phosphate/phosphate antiporter, GlpT, and lactose transporter, LacY, has recently been determined. On the basis of extensive prediction and fold recognition analyses (at the MetaServer), GlpT was proposed as the best structural template on which the arrangement of TM segments of the Pho84 transporter was fit, using the comparative structural modeling program MODELLER. To initiate an evaluation of the appropriateness of the Pho84 model, we have performed two direct tests by targeting spin labels to putative TM segments 8 and 12. Electron paramagnetic resonance spectroscopy was then applied on purified and spin labeled Pho84. The line shape from labels located at both positions is consistent with the structural environment predicted by the template-generated model, thus supporting the model.

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published
keywords
Amino Acid Sequence, Blotting, Western, Chromatography, Affinity, Electron Spin Resonance Spectroscopy, Electrophoresis, Polyacrylamide Gel, Escherichia coli, Fungal Proteins, Models, Molecular, Models, Structural, Molecular Sequence Data, Mutagenesis, Site-Directed, Protein Conformation, Protein Folding, Protein Structure, Secondary, Protein Structure, Tertiary, Proton-Phosphate Symporters, Recombinant Fusion Proteins, Saccharomyces cerevisiae Proteins, Sequence Homology, Amino Acid, Spin Labels, Journal Article, Research Support, Non-U.S. Gov't
in
FEBS Letters
volume
578
issue
3
pages
8 - 262
publisher
Wiley-Blackwell
external identifiers
  • scopus:10044247238
ISSN
0014-5793
DOI
10.1016/j.febslet.2004.11.012
language
English
LU publication?
no
id
2a6e791f-90e5-4b9f-b050-89a2d379bb00
date added to LUP
2017-10-19 20:11:22
date last changed
2017-10-23 09:50:27
@article{2a6e791f-90e5-4b9f-b050-89a2d379bb00,
  abstract     = {<p>The phosphate transporter Pho84 of Saccharomyces cerevisiae is predicted to contain 12 transmembrane (TM) regions, divided into two partially duplicated parts of 6 TM segments. The three-dimensional (3D) organization of the Pho84 protein has not yet been determined. However, the 3D crystal structure of the Escherichia coli MFS glycerol-3-phosphate/phosphate antiporter, GlpT, and lactose transporter, LacY, has recently been determined. On the basis of extensive prediction and fold recognition analyses (at the MetaServer), GlpT was proposed as the best structural template on which the arrangement of TM segments of the Pho84 transporter was fit, using the comparative structural modeling program MODELLER. To initiate an evaluation of the appropriateness of the Pho84 model, we have performed two direct tests by targeting spin labels to putative TM segments 8 and 12. Electron paramagnetic resonance spectroscopy was then applied on purified and spin labeled Pho84. The line shape from labels located at both positions is consistent with the structural environment predicted by the template-generated model, thus supporting the model.</p>},
  author       = {Lagerstedt, Jens O and Voss, John C and Wieslander, Ake and Persson, Bengt L},
  issn         = {0014-5793},
  keyword      = {Amino Acid Sequence,Blotting, Western,Chromatography, Affinity,Electron Spin Resonance Spectroscopy,Electrophoresis, Polyacrylamide Gel,Escherichia coli,Fungal Proteins,Models, Molecular,Models, Structural,Molecular Sequence Data,Mutagenesis, Site-Directed,Protein Conformation,Protein Folding,Protein Structure, Secondary,Protein Structure, Tertiary,Proton-Phosphate Symporters,Recombinant Fusion Proteins,Saccharomyces cerevisiae Proteins,Sequence Homology, Amino Acid,Spin Labels,Journal Article,Research Support, Non-U.S. Gov't},
  language     = {eng},
  month        = {12},
  number       = {3},
  pages        = {8--262},
  publisher    = {Wiley-Blackwell},
  series       = {FEBS Letters},
  title        = {Structural modeling of dual-affinity purified Pho84 phosphate transporter},
  url          = {http://dx.doi.org/10.1016/j.febslet.2004.11.012},
  volume       = {578},
  year         = {2004},
}