asymmetric switching in a homodimeric abc transporter a simulation study不对称转换homodimeric abc转运蛋白模拟研究.pdfVIP

Asymmetric Switching in a Homodimeric ABC Transporter: A Simulation Study 1,2 2 2 1 Jussi Aittoniemi , Heidi de Wet , Frances M. Ashcroft , Mark S. P. Sansom * 1 Department of Biochemistry, University of Oxford, Oxford, United Kingdom, 2 Department of Physiology, Anatomy Genetics, University of Oxford, Oxford, United Kingdom Abstract ABC transporters are a large family of membrane proteins involved in a variety of cellular processes, including multidrug and tumor resistance and ion channel regulation. Advances in the structural and functional understanding of ABC transporters have revealed that hydrolysis at the two canonical nucleotide-binding sites (NBSs) is co-operative and non-simultaneous. A conserved core architecture of bacterial and eukaryotic ABC exporters has been established, as exemplified by the crystal structure of the homodimeric multidrug exporter Sav1866. Currently, it is unclear how sequential ATP hydrolysis arises in a symmetric homodimeric transporter, since it implies at least transient asymmetry at the NBSs. We show by molecular dynamics simulation that the initially symmetric structure of Sav1866 readily undergoes asymmetric transitions at its NBSs in a pre-hydrolytic nucleotide configuration. MgATP-binding residues and a network of charged residues at the dimer interface are shown to form a sequence of putative molecular switches that allow ATP hydrolysis only at one NBS. We extend our findings to eukaryotic ABC exporters which often consist of two non-identical half-transporters, frequently with degeneracy substitutions at one of their two NBSs. Interestingly, many residues involved in asymmetric conformational switching in Sav1866 are substituted in degenerate eukaryotic NBS. This finding strengthens