Abstract

Multidrug-resistance-associated protein 6 (MRP6/ABCC6) is a protein belonging to the ABC transporter family. Proteins in this family share many characteristic structural features, including two membrane-spanning domains and two nucleotide-binding domains (NBD1 and NBD2), that function cooperatively but not equally bind and hydrolyze ATP. The MRP6 protein is structurally and functionally poorly characterized. Previously, we showed, by NMR spectroscopy, that a fragment of MRP6-NBD1 presents helical structure and fluorescence experiments demonstrated that peptide binds ATP. These data suggested that the study on selected regions could be a valid approach to define the structure of MRP6. In the present study, to better characterize MRP6-NBD1, we report data of CD spectroscopy, nucleotide binding and ATP hydrolysis on two different polypeptides, one corresponding to the full-length NBD1 domain (residues from Asp-627 to Leu-851) and the oth We report that both polypeptides are highly structured in aqueous buffer and in 20% trifluoroethanol showing considerable helical and β-structure content. The ATP hydrolysis activity is exhibited only by the full-length NBD1 domain. Comparison between our

Keywords: ABC proteins, ATP binding and hydrolysis, CD spectroscopy, fluorescence spectroscopy, MRP6/ABCC6 (Multidrug Resistance Protein 6), nucleotide binding domain 1, Human Transporter Protein MRP6, Multidrug-resistance-associated protein 6, MRP6/ABCC6, ABC transporter, NBD1, NBD2, NMR spectroscopy, MRP6-NBD1, nucleotide binding, ATP hydrolysis, polypeptides, Arg-648, Thr-805, trifluoroethanol, nucleotide, extracytosolic N-terminus, Pseudoxanthoma elasticum, E748-A785, Sav1866, PXE-associated missense mutations, nucleotide sandwich dimer, circular dichroism (CD) spectroscopic study, PCR Amplification, THERMOHYBAID PCR System, DNA sequencing, Luria Bertani (LB) plates, Monoclonal Anti-polyHistidine-Peroxidase antibody, bovine serum albumin, imidazole, Fluorog-3 spectrometer, SIGMAPLOT 11 software, gluthatione S-Transferase