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- W2048066411 abstract "Bacterial ATP-Binding Cassette (ABC) transporters are vital for nutrient uptake. Structural and functional studies have revealed two distinct types (I and II) of importers based primarily upon mechanistic differences. The ribose transporter (RbsABC) in E. coli is a tripartite ABC importer. RbsABC consists of a cytoplasmic ATP-binding cassette, RbsA, with dual fused nucleotide-binding domains (NBD), a homodimeric transmembrane domain (TMD), RbsC, and a periplasmic substrate binding protein (SBP), RbsB. This study, using Electron Paramagnetic Resonance (EPR) spectroscopy and biochemical methods, demonstrates that RbsABC is a non-canonical ABC transporter that shares some structural and functional features with both type I and type II importers. ATP hydrolysis in the NBD is stimulated by the presence of substrate-loaded SBP, leading to ribose release, which is typical of type I importers. Quantitative measurements obtained from site-directed spin-labeling EPR spectroscopy reveal that hydrolysis promotes the opening of RbsB to release ribose and subsequently RbsB. Post-hydrolysis, both nucleotide and magnesium are required to maintain the complex interactions of RbsC and RbsA, based upon co-purification studies and EPR experiments, which is atypical for ABC importers of either type, where the high-affinity TMD-NBD association is maintained perpetually. In addition, the presence of ribose has a destabilizing effect on RbsB-RbsC interactions, a type II importer behavior. Further, RbsB and RbsC interact in an unusual apo-complex consisting only of the SBP-TMD. Finally, substrate transport is accomplished by hydrolysis at a single consensus site in RbsA, with the second site rendered degenerate by mutations of conserved amino acids. This feature is not uncommon to ABC exporters, but heretofore unobserved in importers. Taking these observations together, RbsABC appears to transport substrate by a distinct mechanism, and offers an opportunity to gain insight into the function of non-canonical systems." @default.
- W2048066411 created "2016-06-24" @default.
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- W2048066411 date "2015-01-01" @default.
- W2048066411 modified "2023-09-25" @default.
- W2048066411 title "Conformational Changes and Complex Formation of the Non-Canonical Ribose ABC Transporter" @default.
- W2048066411 doi "https://doi.org/10.1016/j.bpj.2014.11.802" @default.
- W2048066411 hasPublicationYear "2015" @default.
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