ABSTRACT
ABO3-type perovskite oxides have been regarded as a kind of potential catalyst for peroxymonosulfate (PMS) activation. But some limitations such as specific pH conditions and coexisting ion interference restrict its practical application. Herein, a lanthanum copper Ruddlesden-Popper perovskite oxide (La2CuO4) was successfully synthesized through the sol-gel process and applied in the activation of PMS. And for the first time the La2CuO4/PMS system was used for tetracycline hydrochloride (TC-HCl) degradation. Results showed that La2CuO4 was a potential PMS activation catalyst in the removal of antibiotics. At optimized condition (0.2 g/L catalysts, 1 mM PMS, pH0 6.9), 96.05% of TC-HCl was removed in 30 min. In experiments of debugging control conditions, over a wide pH range of 3-11, more than 90% of TC-HCl can be removed. In the natural water treatment process, TC-HCl removal rates of about 84.2% and 70.3% were obtained in tap water and River water, respectively. According to the reusability and stability tests and the results of FTIR and XPS analysis, La2CuO4 had high structural and chemical stability. Electron paramagnetic resonance (EPR) suggested that the active species including ·OH, SO4-· and 1O2 were detected in degradation reaction. Finally, reasonable reaction mechanisms and possible degradation pathways of TC-HCl were proposed. These results indicate that La2CuO4 can act as a potential catalyst for PMS activation to degrade TC-HCl in water.
Subject(s)
Copper , Tetracycline , Lanthanum , Oxides , Peroxides/chemistryABSTRACT
Usher syndrome (USH) is the leading cause of hereditary hearing-vision loss in humans. PDZ domain-containing 7 (PDZD7) has been reported to be a modifier of and contributor to USH. PDZD7 co-localizes with USH2 proteins in the inner ear hair cells and is essential for ankle-link formation and stereocilia development. PDZD7 contains three PDZ domains and a low-complexity region between the last two PDZ domains, which has been overlooked in the previous studies. Here we characterized a well-folded harmonin homology domain (HHD) from the middle region and solved the PDZD7 HHD structure at the resolution of 1.49 Å. PDZD7 HHD adopts the same five-helix fold as other HHDs found in Harmonin and Whirlin; however, in PDZD7 HHD, a unique α1N helix occupies the canonical binding pocket, suggesting a distinct binding mode. Moreover, we found that the PDZD7 HHD domain can bind lipid and mediate the localization of PDZD7 to the plasma membrane in HEK293T cells. Intriguingly, a hearing-loss mutation at the N-terminal extension region of the HHD can disrupt the lipid-binding ability of PDZD7 HHD, suggesting that HHD-mediated membrane targeting is required for the hearing process. This structural and biochemical characterization of the PDZD7 HHD region provides mechanistic explanations for human deafness-causing mutations in PDZD7. Furthermore, this structure will also facilitate biochemical and functional studies of other HHDs.
ABSTRACT
Enzymes or enzyme complexes can be concentrated in different cellular loci to modulate distinct functional processes in response to specific signals. How cells condense and compartmentalize enzyme complexes for spatiotemporally distinct cellular events is not well understood. Here we discover that specific and tight association of GIT1 and ß-Pix, a pair of GTPase regulatory enzymes, leads to phase separation of the complex without additional scaffolding molecules. GIT1/ß-Pix condensates are modular in nature and can be positioned at distinct cellular compartments, such as neuronal synapses, focal adhesions, and cell-cell junctions, by upstream adaptors. Guided by the structure of the GIT/PIX complex, we specifically probed the role of phase separation of the enzyme complex in cell migration and synapse formation. Our study suggests that formation of modular enzyme complex condensates via phase separation can dynamically concentrate limited quantities of enzymes to distinct cellular compartments for specific and optimal signaling.
Subject(s)
Cell Cycle Proteins/metabolism , GTPase-Activating Proteins/metabolism , Rho Guanine Nucleotide Exchange Factors/metabolism , Signal Transduction , Animals , Cell Cycle Proteins/chemistry , GTPase-Activating Proteins/chemistry , HEK293 Cells , HeLa Cells , Humans , Mice , Models, Molecular , Paxillin/metabolism , Protein Binding , Recombinant Proteins/metabolismABSTRACT
The PSD-95 family of membrane-associated guanylate kinases (MAGUKs) are major synaptic scaffold proteins and play crucial roles in the dynamic regulation of dendritic remodelling, which is understood to be the foundation of synaptogenesis and synaptic plasticity. The guanylate kinase (GK) domain of MAGUK family proteins functions as a phosphor-peptide binding module. However, the GK domain of PSD-95 has been found to directly bind to a peptide sequence within the C-terminal region of neuronal-specific microtubule-associated protein 1A (MAP1A), although the detailed molecular mechanism governing this phosphorylation-independent interaction at the atomic level is missing. In the present study, we determine the crystal structure of PSD-95 GK in complex with the MAP1A peptide at 2.6-Å resolution. The complex structure reveals that, unlike a linear and elongated conformation in the phosphor-peptide/GK complexes, the MAP1A peptide adopts a unique conformation with a stretch of hydrophobic residues far from each other in the primary sequence clustering and interacting with the 'hydrophobic site' of PSD-95 GK and a highly conserved aspartic acid of MAP1A (D2117) mimicking the phosphor-serine/threonine in binding to the 'phosphor-site' of PSD-95 GK. We demonstrate that the MAP1A peptide may undergo a conformational transition upon binding to PSD-95 GK. Further structural comparison of known DLG GK-mediated complexes reveals the target recognition specificity and versatility of DLG GKs.
Subject(s)
Intracellular Signaling Peptides and Proteins/chemistry , Membrane Proteins/chemistry , Microtubule-Associated Proteins/chemistry , Phosphoserine/chemistry , Phosphothreonine/chemistry , Recombinant Fusion Proteins/chemistry , Amino Acid Sequence , Animals , Binding Sites , Cloning, Molecular , Crystallography, X-Ray , Disks Large Homolog 4 Protein , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression , Glutathione Transferase/chemistry , Glutathione Transferase/genetics , Glutathione Transferase/metabolism , HEK293 Cells , Humans , Hydrophobic and Hydrophilic Interactions , Intracellular Signaling Peptides and Proteins/genetics , Intracellular Signaling Peptides and Proteins/metabolism , Membrane Proteins/genetics , Membrane Proteins/metabolism , Mice , Microtubule-Associated Proteins/genetics , Microtubule-Associated Proteins/metabolism , Models, Molecular , Molecular Mimicry , Phosphoserine/metabolism , Phosphothreonine/metabolism , Protein Binding , Protein Interaction Domains and Motifs , Protein Structure, Secondary , Rats , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Sequence Alignment , Sequence Homology, Amino AcidABSTRACT
The membrane-associated guanylate kinase (MAGUK) scaffold proteins share a signature guanylate kinase (GK) domain. Despite their diverse functional roles in cell polarity control and synaptic signaling, the currently known mode of action of MAGUK GK is via its binding to phosphorylated short peptides from target proteins. Here, we discover that the GK domain of DLG MAGUK binds to an unphosphorylated and autonomously folded domain within the stalk region (MAGUK binding stalk [MBS] domain) of a kinesin motor KIF13B with high specificity and affinity. The structure of DLG4 GK in complex with KIF13B MBS reveals the molecular mechanism governing this atypical GK/target recognition mode and provides insights into DLG/KIF13B complex-mediated regulation of diverse cellular processes such as asymmetric cell division. We further show that binding to non-phosphorylated targets is another general property of MAGUK GKs, thus expanding the mechanisms of action of the MAGUK family proteins.
Subject(s)
Kinesins/chemistry , Kinesins/metabolism , Membrane Proteins/chemistry , Membrane Proteins/metabolism , Binding Sites , Humans , Models, Molecular , Phosphorylation , Protein Binding , Protein Domains , Protein Folding , Protein Structure, SecondaryABSTRACT
The tumor suppressors Discs Large (Dlg), Lethal giant larvae (Lgl) and Scribble are essential for the establishment and maintenance of epithelial cell polarity in metazoan. Dlg, Lgl and Scribble are known to interact strongly with each other genetically and form the evolutionarily conserved Scribble complex. Despite more than a decade of extensive research, it has not been demonstrated whether Dlg, Lgl and Scribble physically interact with each other. Here, we show that Dlg directly interacts with Lgl in a phosphorylation-dependent manner. Phosphorylation of any one of the three conserved Ser residues situated in the central linker region of Lgl is sufficient for its binding to the Dlg guanylate kinase (GK) domain. The crystal structures of the Dlg4 GK domain in complex with two phosphor-Lgl2 peptides reveal the molecular mechanism underlying the specific and phosphorylation-dependent Dlg/Lgl complex formation. In addition to providing a mechanistic basis underlying the regulated formation of the Scribble complex, the structure of the Dlg/Lgl complex may also serve as a starting point for designing specific Dlg inhibitors for targeting the Scribble complex formation.
