The nucleotide-binding domain 2 of the human transporter protein MRP6

  • Angela Ostuni
  • Rocchina Miglionico
  • Magnus Monné
  • Maria Antonietta Castiglione Morelli
  • Faustino Bisaccia
Article

Abstract

Multidrug-resistance-associated protein 6 (MRP6/ABCC6) belongs to the ABC transporter family, whose members share many characteristic features including membrane domains and two nucleotide-binding domains (NBD1 and NBD2). These function cooperatively to bind and hydrolyze ATP for the transport of substrates across biological membranes. In this study, MRP6-NBD2 (residues 1252–1503) was expressed in Escherichia coli, purified and structurally and functionally characterized. CD spectra suggested that the protein is folded. Furthermore, NBD2 is shown to be biologically active as it binds ATP and presents ATPase activity although significantly lower compared with isolated NBD1. The mixture of NBD2 and NBD1 exhibited an activity similar to the NBD2 alone, indicating that NBD1 and NBD2 form a heterodimer with the latter limiting ATP hydrolysis. These findings suggest that NBD1 has a higher tendency to form an active homodimer, which is also supported by in silico analysis of energy-minimized dimers of the homology models of the two domains.

Keywords

ABC proteins ATP binding and hydrolysis CD spectroscopy Fluorescence spectroscopy MRP6/ABCC6 (Multidrug Resistance Protein 6) Nucleotide binding domain 2 In silico analysis 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Angela Ostuni
    • 1
  • Rocchina Miglionico
    • 1
  • Magnus Monné
    • 1
  • Maria Antonietta Castiglione Morelli
    • 1
  • Faustino Bisaccia
    • 1
  1. 1.Department of Chemistry “Antonio Mario Tamburro”University of BasilicataPotenzaItaly

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