Sökning: onr:"swepub:oai:DiVA.org:liu-58961" >
Modeling and Mutati...
Modeling and Mutational analysis of Anion transporter 1 protein of Arabidopsis thaliana
-
- Ruiz-Pavon, L (författare)
- Linnaeus University, School of Natural Sciences, Kalmar, Sweden,
-
- Karlsson, Patrik (författare)
- Linköpings universitet,Molekylär genetik,Tekniska högskolan
-
- Carlsson, Jonas (författare)
- Linköpings universitet,Bioinformatik,Tekniska högskolan
-
visa fler...
-
- Samyn, D (författare)
- Linnaeus University, School of Natural Sciences, Kalmar, Sweden,
-
- Persson, Bengt (författare)
- Linköpings universitet,Bioinformatik,Tekniska högskolan
-
- Persson, B L (författare)
- Linnaeus University, School of Natural Sciences, Kalmar, Sweden,
-
- Spetea Wiklund, Cornelia (författare)
- Linköpings universitet,Molekylär genetik,Tekniska högskolan
-
visa färre...
-
(creator_code:org_t)
- Wiley-Blackwell, 2010
- 2010
- Engelska.
-
Ingår i: The FEBS Journal. - : Wiley-Blackwell. - 1742-464X .- 1742-4658. ; 277:Suppl. 1, s. 231-231
- Relaterad länk:
-
https://urn.kb.se/re...
Abstract
Ämnesord
Stäng
- The thylakoid anion transporter 1 (ANTR1) from Arabidopsisthaliana, has been characterized as a Na-dependent Pi transporter when expressed in E. coli (1), but no data is yet available for the protein structure and amino acids involved in transport of Pi. In this study a three-dimensional structural model of ANTR1 was constructed in silico using the crystal structure of glycerol-3- phosphate/phosphate antiporter from E. coli as a template. Based on Multiple Sequence Alignments (MSAs) with other plant ANT- Rs and mammalian SLC17 homologues, five highly conserved amino acids involved in Pi transport have been identified, namely Arg-120, Ser-124 and Arg-201 inside the putative translocation pathway, Arg-228 and Asp-382 exposed at the cytoplasmic sur- face of the protein. The activity of the protein as a Na-dependent Pi transporter in the wild type and mutants was analyzed by het- erologous expression and uptake of radioactive Pi into E. coli cells. Substitution of the three Arg (120, 201 and 228) for Glu residues and of Asp-382 for an Asn residue resulted in an inac- tive ANTR1 transporter. All other mutants had sufficient activity to allow measurement of kinetic parameters, attesting that the mutated proteins were functional. Based on our results, we pro- pose that Arg-201 is a critical residue for substrate binding and translocation, whereas Ser-124 may function as periplasmic gate- way for Na+ ions. Residue Arg-120 plays an important role in Pi binding and associated conformational changes, and finally that Arg-228 and Asp-382 only weakly participate in interactions allowing conformational changes to occur at the cytoplasmic sur-face of the transporter.
Nyckelord
- TECHNOLOGY
- TEKNIKVETENSKAP
Publikations- och innehållstyp
- vet (ämneskategori)
- art (ämneskategori)
Hitta via bibliotek
Till lärosätets databas