lnu.sePublications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Functionally important amino acids in the Arabidopsis thylakoid phosphate transporter: Homology modeling and site-directed mutagenesis
Division of Molecular Genetics, Department of Physics, Chemistry and Biology, Linköping University and School of Natural Sciences, Linnaeus university.
Division of Molecular Genetics, Department of Physics, Chemistry and Biology, Linköping University .
Division of Bioinformatics, Department of Physics, Chemistry and Biology, Linköping University .
Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.ORCID iD: 0000-0002-0381-251X
Show others and affiliations
2010 (English)In: Biochemistry, ISSN 0006-2960, E-ISSN 1520-4995, Vol. 49, no 30, 6430-6439 p.Article in journal (Refereed) Published
Abstract [en]

The anion transporter 1 (ANTR1) from Arabidopsis thaliana, homologous to the mammalian members of the solute carrier 17 (SLC17) family, is located in the chloroplast thylakoid membrane. When expressed heterologously in Escherichia coli, ANTR1 mediates a Na+-dependent active transport of inorganic phosphate (Pi). The aim of this study was to identify amino acid residues involved in Pi binding and translocation by ANTR1 and in the Na+ dependence of its activity. A three-dimensional structural model of ANTR1 was constructed using the crystal structure of glycerol 3-phosphate/phosphate antiporter from E. coli as a template. Based on this model and multiple sequence alignments, five highly conserved residues in plant ANTRs and mammalian SLC17 homologues have been selected for site-directed mutagenesis, namely, Arg-120, Ser-124, and Arg-201 inside the putative translocation pathway and Arg-228 and Asp-382 exposed at the cytoplasmic surface of the protein. The activities of the wild-type and mutant proteins have been analyzed using expression in E. coli and radioactive Pi transport assays and compared with bacterial cells carrying an empty plasmid. The results from Pi- and Na+-dependent kinetics indicate the following: (i) Arg-120 and Arg-201 may be important for binding and translocation of the substrate; (ii) Ser-124 may function as a transient binding site for Na+ ions in close proximity to the periplasmic side; (iii) Arg-228 and Asp-382 may participate in interactions associated with protein conformational changes required for full transport activity. Functional characterization of ANTR1 should provide useful insights into the function of other plant and mammalian SLC17 homologous transporters.

Place, publisher, year, edition, pages
American Chemical Society , 2010. Vol. 49, no 30, 6430-6439 p.
Keyword [en]
Antr1, phosphate transport, Arabidopsis, Major facilitator superfamily, Escherichia coli, Lactose permease, Inorganic-phosphate, transmembrane topology, Membrane-proteins, Crystal structure, Envelope, Family, Cotransporter
National Category
Biochemistry and Molecular Biology
Research subject
Chemistry, Biochemistry
Identifiers
URN: urn:nbn:se:lnu:diva-6914DOI: 10.1021/bi100239jOAI: oai:DiVA.org:lnu-6914DiVA: diva2:332025
Projects
Characterization of sensors and signal transduction in regulation of phosphate uptake systems.
Funder
Swedish Research Council, 621-2007-6144
Available from: 2010-07-30 Created: 2010-07-30 Last updated: 2017-02-10Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full text

Search in DiVA

By author/editor
Ruiz-Pavon, LorenaSamyn, Dieter R.Persson, Bengt L.
By organisation
School of Natural Sciences
In the same journal
Biochemistry
Biochemistry and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar

Altmetric score

Total: 91 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf