The Dynamic Multisite Interactions between Two Intrinsically Disordered Proteins.

Protein interactions involving intrinsically disordered proteins (IDPs) comprise a variety of binding modes, from the well-characterized folding upon binding to dynamic fuzzy complexes. To date, most studies concern the binding of an IDP to a structured protein, while the interaction between two IDPs is poorly understood. In this study, NMR, smFRET, and molecular dynamics (MD) simulation are combined to characterize the interaction between two IDPs, the C-terminal domain (CTD) of protein 4.1G and the nuclear mitotic apparatus (NuMA) protein. It is revealed that CTD and NuMA form a fuzzy complex with remaining structural disorder. Multiple binding sites on both proteins were identified by molecular dynamics and mutagenesis studies. This study provides an atomic scenario in which two IDPs bearing multiple binding sites interact with each other in dynamic equilibrium. The combined approach employed here could be widely applicable for investigating IDPs and their dynamic interactions.

An endoparasitoid Cretaceous fly and the evolution of parasitoidism.

Parasitoidism is a key innovation in insect evolution, and parasitoid insects, nowadays, play a significant role in structuring ecological communities. Despite their diversity and ecological impact, little is known about the early evolution and ecology of parasitoid insects, especially parasitoid true flies (Diptera). Here, we describe a bizarre fly, Zhenia xiai gen. et sp. nov., from Late Cretaceous Burmese amber (about 99 million years old) that represents the latest occurrence of the family Eremochaetidae. Z. xiai is an endoparasitoid insect as evidenced by a highly developed, hypodermic-like ovipositor formed by abdominal tergites VIII + IX that was used for injecting eggs into hosts and enlarged tridactylous claws supposedly for clasping hosts. Our results suggest that eremochaetids are among the earliest definite records of parasitoid insects. Our findings reveal an unexpected morphological specialization of flies and broaden our understanding of the evolution and diversity of ancient parasitoid insects.

Conformational Dynamics of apo-GlnBP Revealed by Experimental and Computational Analysis.

The glutamine binding protein (GlnBP) binds l-glutamine and cooperates with its cognate transporters during glutamine uptake. Crystal structure analysis has revealed an open and a closed conformation for apo- and holo-GlnBP, respectively. However, the detailed conformational dynamics have remained unclear. Herein, we combined NMR spectroscopy, MD simulations, and single-molecule FRET techniques to decipher the conformational dynamics of apo-GlnBP. The NMR residual dipolar couplings of apo-GlnBP were in good agreement with a MD-derived structure ensemble consisting of four metastable states. The open and closed conformations are the two major states. This four-state model was further validated by smFRET experiments and suggests the conformational selection mechanism in ligand recognition of GlnBP.

Molecular basis of the general base catalysis of an α/ß-hydrolase catalytic triad.

The serine-histidine-aspartate triad is well known for its covalent, nucleophilic catalysis in a diverse array of enzymatic transformations. Here we show that its nucleophilicity is shielded and its catalytic role is limited to being a specific general base by an open-closed conformational change in the catalysis of (1R,6R)-2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate synthase (or MenH), a typical α/ß-hydrolase fold enzyme in the vitamin K biosynthetic pathway. This enzyme is found to adopt an open conformation without a functional triad in its ligand-free form and a closed conformation with a fully functional catalytic triad in the presence of its reaction product. The open-to-closed conformational transition involves movement of half of the α-helical cap domain, which causes extensive structural changes in the α/ß-domain and forces the side chain of the triad histidine to adopt an energetically disfavored gauche conformation to form the functional triad. NMR analysis shows that the inactive open conformation without a triad prevails in ligand-free solution and is converted to the closed conformation with a properly formed triad by the reaction product. Mutation of the residues crucial to this open-closed transition either greatly decreases or completely eliminates the enzyme activity, supporting an important catalytic role for the structural change. These findings suggest that the open-closed conformational change tightly couples formation of the catalytic triad to substrate binding to enhance the substrate specificities and simultaneously shield the nucleophilicity of the triad, thus allowing it to expand its catalytic power beyond the nucleophilic catalysis.

How does population aging affect carbon emissions?-analysis based on the multiple mediation effect model.

This paper explores the effect mechanism of population aging on carbon dioxide emissions using provincial panel data in China from 2005 to 2019 and examines how aging affects carbon dioxide emissions through stepwise regression test and Sobel test respectively by constructing a multiple mediating effect model. The following conclusions have been drawn: (1) the results of the baseline regression show that population aging has a significant inhibitory effect on carbon emissions. (2) The results of intermediary effect test show that the intermediary effect of population aging through technological progress, industrial structure, consumption level, and consumption structure is significant. (3) The results of heterogeneity test show that the mediating effect of technological progress is not significant in the eastern region, central region, and western region, and the mediating effect of industrial structure and consumption structure is significant in the eastern region and central region, and the mediating effect of consumption level is significant in the eastern region and western region. (4) The results of the robustness test also verify the mediating effect conclusion once again and prove the robustness of the results. Overall, as an aging society country, China should take the impact of aging into consideration more in the process of achieving emission peak and carbon neutrality.

Ligand-bound glutamine binding protein assumes multiple metastable binding sites with different binding affinities.

Protein dynamics plays key roles in ligand binding. However, the microscopic description of conformational dynamics-coupled ligand binding remains a challenge. In this study, we integrate molecular dynamics simulations, Markov state model (MSM) analysis and experimental methods to characterize the conformational dynamics of ligand-bound glutamine binding protein (GlnBP). We show that ligand-bound GlnBP has high conformational flexibility and additional metastable binding sites, presenting a more complex energy landscape than the scenario in the absence of ligand. The diverse conformations of GlnBP demonstrate different binding affinities and entail complex transition kinetics, implicating a concerted ligand binding mechanism. Single molecule fluorescence resonance energy transfer measurements and mutagenesis experiments are performed to validate our MSM-derived structure ensemble as well as the binding mechanism. Collectively, our study provides deeper insights into the protein dynamics-coupled ligand binding, revealing an intricate regulatory network underlying the apparent binding affinity.