The respiratory complex I transduces
redox energy into an electrochemical
proton gradient in aerobic respiratory chains, powering energy-requiring
processes in the cell. However, despite recently resolved molecular
structures, the mechanism of this gigantic enzyme remains poorly understood.
By combining large-scale quantum and classical simulations with site-directed
mutagenesis and biophysical experiments, we show here how the conformational
state of buried ion-pairs and water molecules control the protonation
dynamics in the membrane domain of complex I and establish evolutionary
conserved long-range coupling elements. We suggest that an electrostatic
wave propagates in forward and reverse directions across the 200 Å
long membrane domain during enzyme turnover, without significant dissipation
of energy. Our findings demonstrate molecular principles that enable
efficient long-range proton–electron coupling (PCET) and how
perturbation of this PCET machinery may lead to development of mitochondrial
disease.
Cyclic peptides have regained interest as potential inhibitors of challenging targets but have often a low bioavailability. The natural product cyclosporine A (CsA) is the textbook exception. Despite its size and polar backbone, it is able to passively cross membranes. This ability is hypothesized to be due to a conformational change from the low-energy conformation in water to a "congruent" conformation that is populated both in water and inside the membrane. Here, we use a combination of NMR measurements and kinetic models based on molecular dynamics simulations to rationalize the difference in the membrane permeability of cyclosporine E (CsE) and CsA. The structure of CsE differs only in a backbone methylation, but its membrane permeability is one order of magnitude lower. The most striking difference is found in the interconversion rates between the conformational states favored in water and in chloroform, which are up to one order of magnitude slower for CsE compared to CsA.
Post-transcriptional mechanisms play a predominant role in the control of microRNA (miRNA) production. Recognition of the terminal loop of precursor miRNAs by RNA-binding proteins (RBPs) influences their processing; however, the mechanistic basis for how levels of individual or subsets of miRNAs are regulated is mostly unexplored. We previously showed that hnRNP A1, an RBP implicated in many aspects of RNA processing, acts as an auxiliary factor that promotes the Microprocessor-mediated processing of pri-mir-18a. Here, by using an integrative structural biology approach, we show that hnRNP A1 forms a 1:1 complex with pri-mir-18a where both RNA recognition motifs (RRMs) bind to cognate RNA sequence motifs in the terminal loop of pri-mir-18a. Terminal loop binding induces an allosteric destabilization of base-pairing in the pri-mir-18a stem that promotes its downstream processing. Our results highlight terminal loop RNA recognition by RBPs as a potential general principle of miRNA biogenesis and regulation.
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