Dataset Information

Conformational Propensities of Peptides Mimicking Transmembrane Helix 5 and Motif C in Wild-type and Mutant Vesicular Acetylcholine Transporters.

ABSTRACT: Vesicular acetylcholine transporter (VAChT) is a member of the major facilitator superfamily (MFS). It contains conserved sequence motifs originally defined in the bacterial multidrug resistance transporter family of the MFS. Motif C (GSLV(227) A(228)PPFGGIL) is located at the C-terminal end of transmembrane helix 5 (TM 5) in VAChT. The motif is rich in glycine and proline residues that often have special roles in backbone conformations of TMs. The A228G mutant of VAChT transports > 3-fold faster than wild type does [Chandrasekaran et al. (2006)J. Neurochem. 98, 1551-1559.]. In the current study, the structure of Loop 4/5, TM 5, and Motif C were taken from a three-dimensional homology model for human VAChT. The peptide was immersed in implicit membrane, energy minimized, and molecular dynamics (MD) were simulated. Kinking and wobbling occur in otherwise helical peptide at the hinge residues L226 and V227. MD also were simulated for A228G single-mutant and V227L-A228A double-mutant peptides to investigate the structural roles of the A228G mutation and beta-branching at V227. Mutant peptides exhibit increased wobbling at the hinge residues, but in the double mutant the increase is less. Because Motif C participates in the interface that mediates hypothesized rocker-switch re-orientation of the acetylcholine binding site during transport, dynamics in Motif C might be an important contributor to transport rate.

SUBMITTER: Luo J

PROVIDER: S-EPMC2882315 | biostudies-literature |

REPOSITORIES: biostudies-literature

ACCESS DATA

Similar Datasets

Project description:Basic architecture of ABC transporters includes two transmembrane domains (TMDs) and two nucleotide-binding domains (NBDs). Although the transport process takes place in the TMDs, which provide the substrate translocation pathway across the cell membrane and control its accessibility between the two sides of the membrane, the energy required for the process is provided by conformational changes induced in the NBDs by binding and hydrolysis of ATP. Nucleotide-dependent conformational changes in the NBDs, therefore, need to be coupled to structural changes in the TMDs. Using molecular dynamics simulations, we have investigated the structural elements involved in the conformational coupling between the NBDs and the TMDs in the Escherichia coli maltose transporter, an ABC importer for which an intact structure is available both in inward-facing and outward-facing conformations. The prevailing model of coupling is primarily based on a single structural motif, known as the coupling helices, as the main structural element for the NBD-TMD coupling. Surprisingly, we find that in the absence of the NBDs the coupling helices can be conformationally decoupled from the rest of the TMDs, despite their covalent connection. That is, the structural integrity of the coupling helices and their tight coupling to the core of the TMDs rely on the contacts provided by the NBDs. Based on the conformational and dynamical analysis of the simulation trajectories, we propose that the core coupling elements in the maltose transporter involve contributions from several structural motifs located at the NBD-TMD interface, namely, the EAA loops from the TMDs, and the Q-loop and the ENI motifs from the NBDs. These three structural motifs in small ABC importers show a high degree of correlation in motion and mediate the necessary conformational coupling between the core of TMDs and the helical subdomains of NBDs. A comprehensive analysis of the structurally known ABC transporters shows a high degree of conservation of the identified 3-motif coupling elements only in the subfamily of small ABC importers, suggesting a distinct mode of NBD-TMD coupling from the other two major ABC transporter folds, namely large ABC importers and ABC exporters.

Dataset Information

Conformational Propensities of Peptides Mimicking Transmembrane Helix 5 and Motif C in Wild-type and Mutant Vesicular Acetylcholine Transporters.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets