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Bis(zinc(II)-dipicolylamine) functionalized sub-2 μm core-shell microspheres for analysis of N-phosphoproteome

ABSTRACT: Herein, bis(zinc(II)-dipicolylamine) functionalized sub-2 μm core-shell silica microspheres (SiO2@DpaZn) were tailored for N-phosphopeptide enrichment at pH 7.7. Contributed by the coordination bonding between Zn(II) and phosphate groups with the KD value of 13.14 μM, N-phosphopeptides could be captured efficiently under neutral conditions. Moreover, contributed by the core-shell structure of SiO2@DpaZn, the fast on-tip enrichment ensured the recovery of N-phosphopeptides up to 44.1%, 3-fold higher than that obtained by in-solution capture. Moreover, 40 determined N-pho sites on 32 substrates were identified in E. coli lysates, among which five had kinase activity. All these results demonstrate that our method is beneficial to promote the discovery of N-phosphoproteins with significant biological functions.

ORGANISM(S): Homo Sapiens

SUBMITTER: Lihua Zhang

PROVIDER: PXD019734 | iProX | Fri Jun 12 00:00:00 BST 2020

REPOSITORIES: iProX

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Bis(zinc(II)-dipicolylamine) functionalized sub-2 μm core-shell microspheres enabled comprehensive analysis of N-phosphoproteome

Project description:Herein, bis(zinc(II)-dipicolylamine) functionalized sub-2 μm core-shell silica microspheres (SiO2@DpaZn) were tailored for N-phosphopeptide enrichment at pH 7.7. Contributed by the coordination bonding between Zn(II) and phosphate groups with the KD value of 13.14 μM, N-phosphopeptides could be captured efficiently under neutral conditions. Moreover, contributed by the core-shell structure of SiO2@DpaZn, the fast on-tip enrichment ensured the recovery of N-phosphopeptides up to 44.1%, 3-fold higher than that obtained by in-solution capture. Moreover, 40 determined N-pho sites on 32 substrates were identified in E. coli lysates, among which five had kinase activity. All these results demonstrate that our method is beneficial to promote the discovery of N-phosphoproteins with significant biological functions.

2020-02-07 | PXD017423 | iProX

Bis(zinc(II)-dipicolylamine) functionalized sub-2 μm core-shell microspheres for analysis of N-phosphoproteome-

2020-08-22 | PXD021067 | iProX

Impact on Arabidopsis thaliana transcriptome of multifunctional silica nanoparticles based on metal atom clusters

Project description:au14-09_silice - clustop transcriptomics. - Understand and categorize plant mechanisms implicated in the interaction with nanoparticles, both in the phenomena responsible for toxicity as in accommodation, see detoxication. - Phytotoxicity of Cs2[Mo6Br14]@SiO2 nanoparticles and their components, i.e. Cs2[Mo6Br14] clusters and SiO2 nanoparticles, has been studied usign Arabidopsis thaliana cell suspension culture. Cs2[Mo6Br14]@SiO2 nanoparticles used here are composed of 7,5% clusters and 92,5% SiO2. Thus, to compare to the impact of a 100 mg/L Cs2[Mo6Br14]@SiO2 nanoparticle treatment (CS), we also treated Arabidopsis cells with clusters at 7,5 mg/L (C), SiO2 at 92,5 mg/L (S), and combined 7,5 mgCMB/L plus 92,5 mgSiO2/L (C+S).

2017-02-01 | GSE93182 | GEO

Nano-silica particles synergistically IgE-mediated mast cell activation

Project description:To deeply investigate the details of the nano-SiO2 effects, we examined the gene expression profile alterations after nano-SiO2 treatment in BMMCs. The difference analysis between the groups showed that 285 genes were significantly expressed after treatment with nano-SiO2. Compared with the blank group, both nano-SiO2 exposure and DNP-HSA stimulation increased the expression of genes related to the MAPK signaling pathway in mast cells to varying degrees.

2022-07-05 | GSE206630 | GEO

Core-shell organization in the mouse claustrum

Project description:Core-shell organization in the mouse claustrum

| PRJNA628885 | ENA

Chiton shell proteome sheds light on stem-group mollusca with biomineralization toolkits

Project description:The appearance of hard mineralized exoskeletons is a critical leap for animal evolution and partially lead to the explosion of diverse animals during the Cambrian, for example, molluscs. A majority of molluscs have mineralized shells to protect themselves. Despite numerous studies that have studied the remarkable mechanical properties of shells, the origin of shell formation is still elusive. Hence, this study investigated the overlooked shell proteome of chitons, which belong to polyplacophoran, Aculifera of Mollusca. By comparing the shell proteome to well-studied Conchifera groups, we inferred possible ancestral biomineralization toolkits of stem-group Mollusca. Taking advantage of the recently sequenced chiton mantle transcriptome and genome, eight core biomineralization proteins were identified by proteomics. Surprisingly, in contrast to previous thought that shell formation is convergently evolved, two important shell matrix proteins, Nacrein-like and Pif-like proteins were found to be conserved among Aculifera and Conchifera groups. Our findings identify a missed link of mineralized shell evolution in Mollusca and pose a hypothesis that stem-group molluscs have already evolved core biomineralization toolkits, which likely facilitate the formation of mineralized shells for protection that partially leads to their explosion.

2023-01-03 | PXD033490 | Pride

Global proteomics to study silica nanoparticle-induced cytotoxicity and its mechanisms in HepG2 cells

Project description:Silica nanoparticles (SiO2 NPs) are commonly used in medical and pharmaceutical fields. Research into the cytotoxicity and overall proteomic changes occurring during initial exposure to SiO2 NPs is limited. We investigated the mechanism of toxicity in human liver cells according to exposure time [0, 4, 10, and 16 hours (h)] to SiO2 NPs through proteomic analysis using mass spectrometry. SiO2 NP-induced cytotoxicity through various pathways in HepG2 cells. Interestingly, when cells were exposed to SiO2 NPs for 4 h, the morphology of the cells remained intact, while the expression of proteins involved in mRNA splicing, cell cycle, and mitochondrial function was significantly downregulated. These results show that the toxicity of the nanoparticles affects protein expression even if there is no change in cell morphology at the beginning of exposure to SiO2 NPs. The levels of reactive oxygen species changed significantly after 10 h of exposure to SiO2 NPs, and the expression of proteins associated with oxidative phosphorylation, and the immune system was upregulated. Eventually, these changes in protein expression induced HepG2 cell death. This study provides insights into cytotoxicity evaluation at early stages of exposure to SiO2 NPs through in vitro experiments.

2021-09-10 | PXD024206 | Pride

Gene expression response to silica treatment in wild-type C57 black 6 mice vs Rag1 knockout, NK-depleted mice

Project description:The lung is constantly exposed to potentially pathogenic particles and microorganisms. It has recently become evident that not only innate, but also adaptive immune responses to particulates such as crystalline silica (SiO2) entering the respiratory tract are complex and dynamic events. Although the cellular mechanisms and anatomical consequences involved in the development of silicosis have been extensively studied, they still remain poorly understood. Based on their capacity for immune regulation, lymphocytes may play a role in determining the respiratory response to environmental challenge by SiO2. The objective of this study was to characterize the impact of SiO2 exposure on respiratory immune processes, with particular emphasis on evaluating the importance of lymphocytes in the murine silicosis model. Therefore, we utilized lymphopenic mice including NK deficient, Rag1-/- or a combination (Rag1-/- NK depleted) and demonstrated that SiO2-induced fibrosis and inflammation occur independently of T, B, NK T, and NK cells. Studies in Rag1-/- mice further suggest that lymphocytes may participate in controlling SiO2-induced inflammation through modulation of the Nalp3 inflammasome. This observation may have clinical relevance in the treatment of inflammatory and fibrotic lung diseases that are either refractory or respond sub-optimally to current therapeutics.

2009-12-19 | GSE19563 | GEO

Symmetric Bis-benzimidazoles Are Potent Anti-Staphylococcal Agents with Dual Inhibitory Mechanisms against DNA Gyrase

Project description:Various bis-benzimidazole derivatives have been reported to possess activity against Gram-positive pathogens. No mechanism of action has been elucidated to fully account for the antibacterial activity of this class of compounds. A group of symmetric bis-benzimidazoles (BBZ) designed as anticancer agents have previously been shown to possess moderate antiproliferative activity. We sought to assess the antibacterial activity and mechanism of action of BBZ compounds against Staphylococcus aureus. Antibacterial activities were assessed by determination of minimal inhibitory concentrations (MICs), time-kill curves, and scanning electron microscopy. Transcriptional responses to BBZ treatment were determined using whole genome microarrays. Activities against bacterial type II topoisomerases were investigated using in vitro supercoiling, decatenation, DNA binding, and DNA cleavage inhibition assays. MICs for EMRSA-16 were between 0.03 and 0.5 μg/mL. The compounds showed concentration-dependent bactericidal activity and induced cell swelling and lysis. Transcriptional responses to BBZ were consistent with topoisomerase inhibition and DNA damage. A subset of BBZ compounds inhibited S. aureus DNA gyrase supercoiling activity with IC50 values in the range of 5−10 μM. This inhibition was subsequently shown to operate through both inhibition of binding of DNA gyrase to DNA and accumulation of single-stranded DNA breaks. We conclude that BBZ compounds are potent antistaphylococcal agents and operate at least in part through DNA gyrase inhibition, leading to the accumulation of single-stranded DNA breaks, and by preventing the binding of gyrase to DNA. [Data is also available from http://bugs.sgul.ac.uk/E-BUGS-106]

2012-10-14 | E-BUGS-106 | biostudies-arrayexpress

The microstructural, mineralogical and transcriptional developments of shell biomineralization of Pinctada maxima

Project description:Molluscan larval ontogeny is a highly conserved process typical of 3 principal developmental stages. A characteristic unique to each of these stages is shell design, termed prodissoconch I, prodissoconch II and dissoconch. These shells vary in morphology, mineralogy and microstructure. The discrete temporal transitions in shell biomineralization between these larval stages are utilized in this study to investigate transcriptional involvement in several distinct biomineralization events. Scanning electron microscopy and X-ray diffraction analysis of P. maxima larvae and juveniles collected throughout post-embryonic ontogenesis, document the mineralogy and microstructure of each shelled stage as well as establishing a timeline for transitions in biomineralization. P. maxima larval samples most representative of these biomineralization distinctions and transitions were analyzed for differential gene expression on the microarray platform PmaxArray 1.0. A number of transcripts are reported as differentially expressed in correlation to the mineralization events of P. maxima larval ontogeny. Some of those isolated are known shell matrix genes while others are novel, these are discussed in relation to potential shell formation roles. This interdisciplinary investigation has married the shell developments of P. maxima larval ontogeny with corresponding gene expression profiles, furthering the elucidation of shell biomineralization. Keywords: Temporal expression profiling by array

2010-02-05 | GSE14305 | GEO

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