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 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.
journal_name
FEBS Lettjournal_title
FEBS lettersauthors
Lagerstedt JO,Voss JC,Wieslander A,Persson BLdoi
10.1016/j.febslet.2004.11.012keywords:
subject
Has Abstractpub_date
2004-12-17 00:00:00pages
262-8issue
3eissn
0014-5793issn
1873-3468pii
S0014-5793(04)01376-6journal_volume
578pub_type
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