Analyse eines unkonventionellen Vitamin-Transporters

Abstract

Förster resonance energy transfer (FRET) identifies the co-localization of fluorescent macromolecules on a nanometer scale. FRET between nonidentical and identical fluorophores is called HeteroFRET or HomoFRET, respectively. We applied FRET techniques to analyze the oligomeric state of the core component (BioY) of a bacterial biotin transporter. BioY was fused to fluorescent proteins suitable for Hetero- and HomoFRET analyses. Spectrometric and imaging experiments with living cells strongly support a model with oligomers of BioY as the functional state of the core unit.

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Literatur

  1. [1]

    Rodionov DA, Hebbeln P, Eudes A et al. (2009) A novel class of modular transporters for vitamins in prokaryotes. J Bacteriol 191:42–51

    PubMed  Article  CAS  Google Scholar 

  2. [2]

    Eitinger T, Rodionov DA, Grote M et al. (2011) Canonical and ECF-type ATP-binding cassette importers in prokaryotes: diversity in modular organization and cellular functions. FEMS Microbiol Rev 35:3–67

    PubMed  Article  CAS  Google Scholar 

  3. [3]

    Erkens GB, Majsnerowska M, Ter Beek J et al. (2012) Energy coupling factor-type ABC transporters for vitamin uptake in prokaryotes. Biochemistry 51:4390–4396

    Article  CAS  Google Scholar 

  4. [4]

    Neubauer O, Reiffler C, Behrendt L et al. (2011) Interactions among the A and T units of an ECF-type biotin transporter analyzed by site-specific crosslinking. PLoS One 6:e29087

    PubMed  Article  CAS  Google Scholar 

  5. [5]

    Henderson GB, Zevely EM, Huennekens FM (1979) Mechanism of folate transport in Lactobacillus casei: evidence for a component shared with the thiamine and biotin transport systems. J Bacteriol 137:1308–1314

    PubMed  CAS  Google Scholar 

  6. [6]

    Zhang P, Wang J, Shi Y (2010) Structure and mechanism of the S component of a bacterial ECF transporter. Nature 468:717–720

    PubMed  Article  CAS  Google Scholar 

  7. [7]

    Erkens GB, Berntsson RP, Fulyani F et al. (2011) The structural basis of modularity in ECF-type ABC transporters. Nat Struct Mol Biol 18:755–760

    PubMed  Article  CAS  Google Scholar 

  8. [8]

    Hebbeln P, Rodionov DA, Alfandega A et al. (2007) Biotin uptake in prokaryotes by solute transporters with an optional ATP-binding cassette-containing module. Proc Natl Acad Sci USA 104:2909–2914

    PubMed  Article  Google Scholar 

  9. [9]

    Kirsch F, Frielingsdorf S, Pohlmann A et al. (2012) Essential amino acid residues of BioY reveal that dimers are the functional S unit of the Rhodobacter capsulatus biotin transporter. J Bacteriol 194 (im Druck), doi: 10.1128/JB.00683-12

  10. [10]

    Finkenwirth F, Neubauer O, Gunzenhäuser J et al. (2010) Subunit composition of an energy-coupling-factor-type biotin transporter analysed in living bacteria. Biochem J 431:373–380

    PubMed  CAS  Google Scholar 

  11. [11]

    Grecco HE, Verveer PJ (2011) FRET in cell biology: still shining in the age of super-resolution? ChemPhysChem 12:484–490

    PubMed  Article  CAS  Google Scholar 

  12. [12]

    Bader AN, Hoetzl EG, Hofman J et al. (2011) Homo-FRET imaging as a tool to quantify protein and lipid clustering. ChemPhysChem 12:475–483

    PubMed  Article  CAS  Google Scholar 

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Correspondence to Joanna Ziomkowska or Franziska Kirsch or Andreas Herrmann or Thomas Eitinger.

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Franziska Kirsch, Thomas Eitinger, Joanna Ziomkowska und Andreas Herrmann (v. l. n. r.)

Die Arbeitsgruppe von Andreas Herrmann beschäftigt sich mit Lipid- und Protein-Wechselwirkungen und deren Bedeutung für die Struktur und Dynamik von Membranen. Schwerpunkte in der Arbeitsgruppe von Thomas Eitinger bilden bakterielle Aufnahmesysteme, unter anderem verschiedene Vertreter der ECF-Transporter für Vitamine und Übergangsmetallionen.

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Ziomkowska, J., Kirsch, F., Herrmann, A. et al. Analyse eines unkonventionellen Vitamin-Transporters. Biospektrum 18, 493–496 (2012). https://doi.org/10.1007/s12268-012-0220-x

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