MICROCHEMICAL JOURNAL
·
2023
·
IF 4.8
Yongyin Zhou,Hui Li,Hongjun Luo,Zhexuan Lin,Wenhong Luo
An untargeted UHPLC-MS method was developed for the preliminary screening of differential amino acids between two mouse brain tissue samples, and a targeted UHPLC-MS/MS method was developed for the quantitative determination of amino acids in mouse brain tissue samples. The sample processing procedure, chromatographic conditions and MS parameters were optimized. The mouse brain tissue samples were homogenized with HCl solution (0.01 mol/L), protein precipitated with methanol and centrifugation. For the untargeted method, one sample was derivatized with butanol (D0) and another sample was derivatized with deuterated butanol (D9), then combined together and analyzed with UHPLC-MS. Chromatographic peak area ratio for each amino acid was then used for differential analyses between two samples. Results showed good accuracy with standard spiked samples (83.4–113.0 %). For the targeted quantitative method, the sample supernatants were dried and residues were derivatized with butanol (D0). Internal standards of amino acids derivatized with deuterated butanol (D9) were added before analyzed with UHPLC-MS/MS. The intra- and inter-day precisions were in the intervals of 0.5–8.1 % and 0.3–9.1 %. The LOD and the LOQ were ranged from 0.03 × 10 −3 to 0.92 × 10 −3 µg mL −1 and 0.08 × 10 −3 to 3.08 × 10 −3 µg mL −1 , respectively. Spiked recoveries were from 96.8 % to 111.7 %. Both methods were applied to analyze amino acids in brain tissue of C57BL/6 mice treated and untreated with piperine and the result showed that the levels of glutamic acid, aspartic acid, histidine, asparagine, γ-aminobutyric acid, alanine, leucine and isoleucine significantly decreased in the piperine treated group comparing with the untreated group.
研究领域:
分析化学
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神经生物学
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药理学
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产品关联
DOI
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2023
·Vol. 415
·期 23
·5575-5588
·
IF 4.3
Liu Yuanyuan,Han Yutong,Zhu Wenjie,Luo Qingming,Yuan Jing,Liu Xin
Glycosylation is one of the most common types of post-translational modifications in mammals. It is well known that N-glycans play a key role in cell adhesion, differentiation, synapsis, and myelination during the development of the mammalian central nervous system (CNS). Neuropathological symptoms (such as epilepsy and Alzheimer’s disease) are usually accompanied by N-glycosylation changes. In this study, we extracted N-glycan chains from eight regions of the mouse brain, and combined high-throughput, high-resolution matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) with the Fmoc N-hydroxysuccinimide ester (Fmoc-OSU) derivatization method to improve the sensitivity of glycan detection to characterize the total N-glycans in the mouse brain. A total of 96 N-glycan moieties were detected. An exhaustive examination of the relative abundance of N-glycans, coupled with a comparative analysis of differences, has uncovered discernible variations of statistical significance, including high mannose, fucosylated, sialylated, and galactosylated N-glycans. According to our investigations, a thorough and regionally specific cartography of glycans within the brain can facilitate the investigation of glycan-mediated mechanisms related to both the developmental trajectory and functional output of the brain. Additionally, this approach may serve as a basis for identifying potential biomarkers that are relevant to various brain-associated pathologies. Graphical Abstract
研究领域:
生物化学
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神经科学
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分子生物学
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DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2023
·
IF 4.1
Hongmei Yang,Linxu Wang,Zhaoyang Xie,Simeng Shao,Yi Wu,Weiyin Xu,Bin Gu,Bo Wang
Comprehensive sulfur-nitrosylation (SNO) proteome coverage in complex biological systems remains challenging as a result of the low level of endogenous S-nitrosylation and its chemical instability. Herein, we optimized the synthesis route of SNOTRAP (SNO trapping by triaryl phosphine) probe and the proteomics pipeline (including preventing over-alkylation, sample washing, trypsin digestion). Preventing overalkylation was found to be the key factor resulting in a higher number of identified SNO proteins by evaluating various experimental conditions. With the improved SNOTRAP-based proteomic pipeline, we achieved an improvement of ∼10-fold on identification efficiency, and identified 1181 SNO proteins (1714 SNO sites) in mouse brain, representing the largest repository of endogenous S-nitrosylation. Moreover, we identified the consensus motif of SNO sites, suggesting the correlation with local hydrophobicity, acid-base catalysis, and the surrounding secondary structures for modification of specific cysteines by NO. Collectively, we provide a universal pipeline for the high-coverage identification of low-abundance SNO proteins with high enrichment efficiency, high specificity (98%), good reproducibility, and easy implementation, contributing to the elucidation of the mechanism(s) of nitrosative stress in multiple diseases.
研究领域:
生物化学
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蛋白质组学
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医学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF MASS SPECTROMETRY
·
2022
·Vol. 57
·期 3
·e4813
·
IF 2.394
Liutian Wu,Keke Qi,Minggao Xu,Chengyuan Liu,Yang Pan
Desorption electrospray ionization/post-photoionization (DESI/PI) is a newly developed ionization method by the combination of DESI and post-photoionization for the simultaneous imaging of polar and nonpolar compounds in biological tissues. Dopants are of great importance in DESI/PI for the enhancement of signal intensities through ion–molecule reactions. In this work, to evaluate the performance of dopants in DESI/PI, an efficient homogenate model was developed, and four kinds of dopants (toluene, chlorobenzene, bromobenzene, and anisole) were tested using homogenate of mouse brain tissue as target sample. The influences of the dopants on the signal enhancements of different compounds were explained reasonably by the ionization mechanism. Then, the dopants with their optimum volume contents were applied to the mass spectrometry imaging (MSI). For a comprehensive imaging of various compounds with different polarities, methanol/toluene/formic acid (7:3:0.1) was chosen as the best choice. Finally, the stronger quantitative ability of DESI/PI with toluene as dopant for a few compounds in mouse brain tissue was demonstrated.
研究领域:
分析化学
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生物医学
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产品关联
DOI
PubMed
高影响因子
BIOMEDICAL CHROMATOGRAPHY
·
2019
·Vol. 34
·期 3
·e4767
·
IF 1.748
Yong-hong Liu,Ting-ting Du,Jing-yuan Wan,Hua Zhao,Dan Huang,Jian-sha Li,Xin-hui Jiang
Disorders of certain branched-chain amino acids may be associated with the occurrence and development of non-alcoholic fatty liver disease. Measurement of related branched-chain amino acid levels could provide a reference for the clinical and scientific research of the non-alcoholic fatty liver disease. An established HPLC–FLD method was used to quantify aspartic acid, glutamate, glutamine, glycine, taurine, tyrosine, 4-amino butanoic acid, tryptophan, methionine, valine, phenylalanine, isoleucine and leucine in mouse brain tissue. Brain tissue samples mixed with internal standard (3-aminobutyric acid) were processed, then derivatized with 2- O -phthaldialdehyde, and finally separated on an ODS2 column through gradient elution at a flow rate of 1.0 ml·min −1 . The excitation and emission wavelengths were set at 340 and 455 nm, respectively. The mobile phase A was 100% methanol and the mobile phase B consisted of 30 mmol·L −1 sodium acetate (pH 6.8). The injection volume was 20 μl and the single run time was 45 min. Several parameters, accuracy, precision, and stability, were verified and the results showed the established method had good sensitivity and resolution for all of the 13 compounds and internal standard in mouse brain.
研究领域:
生物化学
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医学研究
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分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYST
·
2022
·Vol. 147
·期 18
·4092-4097
·
IF 5.227
Yumei Xu,Bing Hu,Yanjun Cui,Li Li,Fang Nian,Zhixia Zhang
In this work, we developed a new organic electrochemical probe, 4-(prop-2-yn-1-yloxy)phenol, for the determination of tyrosinase. The designed probe contained a 4-hydroxybenzyloxy moiety for the specific determination of Tyr and a terminal alkyne group for chemical adsorption onto the electrode surface. The oxidation peak of the phenolic group from Pyyp decreased but increased for o-diphenol or o-quinone generated after the reaction between Tyr and Pyyp. The present sensor demonstrated a good linearity with Tyr activity in a dynamic range of 1.0–30 U mL−1. The limit of detection was as low as 0.28 ± 0.12 U mL−1. Moreover, this sensor demonstrated high selectivity for Tyr determination against metal ions, amino acids, ROS and neurotransmitters due to the specific recognition of Tyr. Finally, the developed sensor with high accuracy, high selectivity and long-term stability was successfully applied for evaluating Tyr activity in normal brain homogenate and brain homogenate with Parkinson's disease.
研究领域:
分析化学
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神经科学
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生物化学
✓ 已验证
DOI
PubMed
高影响因子
Journal of Proteomics
·
2023
·
IF 3.3
Yuanyuan Zhang,Yutong Han,Wenjie Zhu,Jing Yuan,Xin Liu
Protein C-termini containing valuable biological information plays a vital role in various physiological processes , such as protein localization, protein recognition, and signal transduction in organisms. However, C-terminal peptide identification is still challenging due to their low abundance and similar physicochemical properties to other digested peptides. Herein, we developed a simple and mild strategy for the enrichment of C-terminal peptides that incorporates selectively 2-pyridinecarbaldehyde (2-PCA) derivatization of α-amine with negative enrichment by NHS resin. Two synthesized peptides were utilized to evaluate the efficiency of 2-PCA derivatization and optimize the coupling conditions of NHS resin. The feasibility of the method was further validated by enriching the C-terminus of the bovine serum albumin (BSA). Finally, this method was successfully applied to the C-terminus analysis of mouse brain tissue, identifying 404 protein C-termini with physicochemical properties unbiasedly. Additionally, the GO and KEGG analyses revealed that these identified proteins are crucial for proper brain function. In summary, our proposed method is effective and has the potential to facilitate comprehensive C-terminal analysis of proteins . Significance Effective enrichment methods are essential for the identification of the proteins C-terminus. In this study, a mild and simple method for negative C-terminal enrichment combined with site-specific derivatization was developed. The enrichment process was simplified and minimized sample loss simultaneously, using 2-PCA derivatization which has high α-amino specificity. Up to 346C-terminal proteins were identified in mouse brain tissue unbiasedly and reliably. This approach has the potential to facilitate comprehensive analysis of protein C-termini in a variety of biological contexts.
研究领域:
生物化学
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蛋白质组学
·
系统生物学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2018
·
IF 6.042
Xiaoqin Zhang,Yuting Zhang,Caiyun Fang,Lei Zhang,Pengyuan Yang,Changchun Wang,Haojie Lu
Palmitoylation, a type of fatty acylation, has vital roles in many biological processes. For ultradeep identification of protein palmitoylation, an enrichment approach based on a novel magnetic microsphere modified with 2,2′-dithiodipyridine (Fe3O4/SiO2-SSPy microsphere) is presented in this study. The Fe3O4/SiO2-SSPy microspheres were synthesized by directly coating thiol-containing silane coupling agent onto the magnetic supraparticles in aqueous solution at room temperature. Due to the intrinsic magnetic properties, high surface-to-volume ratios, and abundant reactive functional groups on the surface, these microspheres enabled direct capture of palmitoylated targets and convenient isolation, contributing to remarkable enrichment selectivity (purifying palmitoylated peptides from mixtures with nonpalmitoylated peptides even at a 1:500 molar ratio) and sensitivity (the detection limit was at femtomole level), thus enabling a global annotation of protein palmitoylation for complex biological samples. We successfully identified 1304 putative palmitoylated proteins from mouse brain tissues by using this method, which is the largest mouse palmitoylome data set to date. Except for those known members, many new proteins and pathways were also found to be regulated by palmitoylation.
研究领域:
生物化学
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蛋白质组学
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纳米技术
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分子生物学
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2021
·
IF 6.057
Jian Zhang,Chengan Guo,Xinming Huo,Xiaoxiao Ma,Xin Li,Zeper Abliz,Yao Chu,Xiaohao Wang,Fei Tang
In recent years, the development of unsaturated lipid isomeric imaging based on the Paternò–Büchi (PB) reaction has improved significantly. However, research on this imaging method in ambient conditions needs to expand. In this paper, a method of PB reaction in the solid phase in ambient conditions is developed, which is combined with air-flow-assisted desorption electrospray ionisation mass spectrometry (AFADESI-MS) to achieve in situ imaging of lipids at an isomeric level. Experiments showed that the efficiency of the PB reaction was much higher when spraying the reagent solution than when sprinkling the reactant powder directly, and it was not lower than that in the liquid phase. This method can simplify the reaction conditions in the imaging process and can be applied to tissue section samples with only 10 min of pre-processing. The study successfully demonstrated the spatial distribution of unsaturated lipid isomers, and the isomeric ratio corresponded to the lesion areas in mouse brain cancer tissues. Due to its simple operation and performance in ambient conditions, this method may be useful for future studies on lipid isomers in tissues.
研究领域:
分析化学
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生物医学工程
·
质谱学
·
脂质组学
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产品关联
DOI
PubMed
高影响因子
APPLIED SURFACE SCIENCE
·
2014
·
IF 2.538
Songbo Xie,Baolin Zhang,Lei Wang,Jun Wang,Xuan Li,Gao Yang,Fabao Gao
PEG and PEG/PEI modified superparamagnetic iron oxide nanoparticles (SPIONs) were synthesized by the thermal decomposition of iron (III) acetylacetonate (Fe(acac) 3 ) in poly (ethylene glycol) (PEG) containing poly (ethylene imine) (PEI) (0 or 0.3 g). PEG/PEI-SPIONs were coated with Tween 80 (PEG/PEI/Tween 80-SPIONs). Fourier transform infrared spectroscopy (FTIR) analyses indicated that PEG , PEG/PEI and PEG/PEI/Tween 80 were attached to the surfaces of the SPIONs. The PEG-SPIONs, PEG/PEI-SPIONs and PEG/PEI/Tween 80-SPIONs performed excellent colloidal stability in the phosphate buffered saline (PBS), and in deionized water with the mean hydrodynamic sizes of 19.5, 21.0, 24.0 nm and the zeta potentials of -5.0, 35.0, 19.0 mV, respectively. All the SPIONs showed low cytotoxicity assessed by the MTT assay. In vivo magnetic resonance imaging (MRI) of the Kunming (KM) mouse brains were performed, the PEG-SPIONs, PEG/PEI-SPIONs and PEG/PEI/Tween 80-SPIONs exhibited vascular imaging effects in bulbus olfactorius, frontal cortex, temporal, thalamus and brain stem of the mouse brains after 24 h intravenous injection of the nanoparticles . The SPIONs have potentials as MRI contrast agents in the mouse brains.
研究领域:
生物医学工程
·
纳米医学
·
影像学
✓ 已验证
DOI
高影响因子
Neuroscience Bulletin
·
2023
·
IF 5.6
Song Changgeng,Zhao Yan,Zhang Jiajia,Dong Ziyi,Kang Xin,Pan Yuqi,Du Jinle,Gao Yiting,Zhang Haifeng,Xi Ye,Ding Hui,Kuang Fang,Wang Wenting,Luo Ceng,Zhang Zhengping,Zhao Qinpeng,Yang Jiazhou,Jiang Wen,Wu Shengxi,Gao Fang
Parvalbumin interneurons belong to the major types of GABAergic interneurons. Although the distribution and pathological alterations of parvalbumin interneuron somata have been widely studied, the distribution and vulnerability of the neurites and fibers extending from parvalbumin interneurons have not been detailly interrogated. Through the Cre recombinase-reporter system, we visualized parvalbumin-positive fibers and thoroughly investigated their spatial distribution in the mouse brain. We found that parvalbumin fibers are widely distributed in the brain with specific morphological characteristics in different regions, among which the cortex and thalamus exhibited the most intense parvalbumin signals. In regions such as the striatum and optic tract, even long-range thick parvalbumin projections were detected. Furthermore, in mouse models of temporal lobe epilepsy and Parkinson’s disease, parvalbumin fibers suffered both massive and subtle morphological alterations. Our study provides an overview of parvalbumin fibers in the brain and emphasizes the potential pathological implications of parvalbumin fiber alterations.
研究领域:
神经科学
·
神经解剖学
·
病理学
·
神经疾病
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2022
·
IF 6.556
Li Zhan,Chengyuan Liu,Keke Qi,Liutian Wu,Ying Xiong,Xuan Zhang,Jianye Zang,Yang Pan
In this work, endogenous metabolites in mouse brain tissue were imaged by negative desorption electrospray ionization (DESI), ammonia assisted DESI (aa-DESI), DESI/post-photoionization (DESI/PI), and ammonia assisted DESI/PI (aa-DESI/PI) mass spectrometry imaging (MSI) strategies. The combined effect of ammonia additive and post-photoionization was found to play an important role in the enhancement of sensitivity and coverage for endogenous analytes under ambient conditions. Compared with DESI, aa-DESI/PI can provide increased signal intensities for metabolites up to 37.1-fold, as well as the imaging of nine more small metabolites ( m/z < 350) (26 for aa-DESI/PI and 17 for DESI) in mouse brain tissue. The results of Pearson correlation analysis and KEGG pathway analysis showed that the enhanced imaging strategy of aa-DESI/PI can facilitate the study of endogenous metabolic pathways and molecular networks. Moreover, the imaging results of mouse tumor tissue demonstrated the promising application of aa-DESI/PI in tumor research.
研究领域:
分析化学
·
生物信息学
·
分子生物学
·
医学影像
✓ 已验证
产品关联
DOI
PubMed
高影响因子
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