Inter-domain Communication Mechanisms in an ABC Importer: A Molecular Dynamics Study of the MalFGK ₂E Complex

Abstract

ATP-Binding Cassette transporters are ubiquitous membrane proteins that convert the energy from ATP-binding and hydrolysis into conformational changes of the transmembrane region to allow the translocation of substrates against their concentration gradient. Despite the large amount of structural and biochemical data available for this family, it is still not clear how the energy obtained from ATP hydrolysis in the ATPase domains is “transmitted” to the transmembrane domains. In this work, we focus our attention on the consequences of hydrolysis and inorganic phosphate exit in the maltose uptake system (MalFGK ₂E) from Escherichia coli. The prime goal is to identify and map the structural changes occurring during an ATP-hydrolytic cycle. For that, we use extensive molecular dynamics simulations to study three potential intermediate states (with 10 replicates each): an ATP-bound, an ADP plus inorganic phosphate-bound and an ADP-bound state. Our results show that the residues presenting major rearrangements are located in the A-loop, in the helical sub-domain, and in the “EAA motif” (especially in the “coupling helices” region). Additionally, in one of the simulations with ADP we were able to observe the opening of the NBD dimer accompanied by the dissociation of ADP from the ABC signature motif, but not from its corresponding P-loop motif. This work, together with several other MD studies, suggests a common communication mechanism both for importers and exporters, in which ATP-hydrolysis induces conformational changes in the helical sub-domain region, in turn transferred to the transmembrane domains via the “coupling helices”.

Author Summary

ABC transporters are membrane proteins that couple ATP binding and hydrolysis with the active transport of substrates across membranes. These transporters form one of the largest families of membrane proteins and they can be found in all phyla of life. Moreover, some members of this family are involved in several genetic diseases (such as cystic fibrosis) and in multidrug resistance in bacteria, fungi and mammals. In this work, we use molecular dynamics simulations to study conformational changes due to ATP hydrolysis in an ABC transporter responsible for maltose uptake in E. coli. These conformational changes arising from one side of the protein (NBDs – Nucleotide Binding domains) where ATP binds, are propagated across the protein to more distant regions. Additionally, we can observe an NBD dimer interface dissociation event upon inorganic phosphate exit. These simulations together with other theoretical studies suggest that there is a general inter-domain communication mechanism common to importers and exporters.