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SWATH-MS Based Secretome Proteomic Analysis of Pseudomonas aeruginosa Against MRSA 基于 SWATH-MS 的铜绿假单胞菌与 MRSA 的分泌组蛋白质组分析。
IF 3.4 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-17 DOI: 10.1002/pmic.202300649
Yi-Feng Zheng, Yu-Sheng Lin, Jing-Wen Huang, Kuo-Tung Tang, Cheng-Yu Kuo, Wei-Chen Wang, Han-Ju Chien, Chih-Jui Chang, Nien-Jen Hu, Chien-Chen Lai

The study uses Sequential Window Acquisition of All Theoretical Fragment Ion Mass Spectra (SWATH)-MS in conjunction with secretome proteomics to identify key proteins that Pseudomonas aeruginosa secretes against methicillin-resistant Staphylococcus aureus (MRSA). Variations in the inhibition zones indicated differences in strain resistance. Multivariate statistical methods were applied to filter the proteomic results, revealing five potential protein biomarkers, including Peptidase M23. Gene ontology (GO) analysis and sequence alignment supported their antibacterial activity. Thus, SWATH-MS provides a comprehensive understanding of the secretome of P. aeruginosa in its action against MRSA, guiding future antibacterial research.

该研究利用序列窗口获取所有理论碎片离子质谱(SWATH)-MS 与分泌组蛋白质组学相结合的方法,确定了铜绿假单胞菌分泌的抗耐甲氧西林金黄色葡萄球菌(MRSA)的关键蛋白质。抑制区的变化表明菌株的耐药性存在差异。应用多元统计方法对蛋白质组结果进行筛选,发现了包括肽酶 M23 在内的五个潜在蛋白质生物标记物。基因本体(GO)分析和序列比对支持了它们的抗菌活性。因此,SWATH-MS 提供了对铜绿假单胞菌抗 MRSA 作用的分泌组的全面了解,为未来的抗菌研究提供了指导。
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引用次数: 0
Contents: Proteomics 20'24 内容:蛋白质组学 20'24
IF 3.4 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-10 DOI: 10.1002/pmic.202470163
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引用次数: 0
Astronaut proteomics: Japan leads the way for transformative studies in space 宇航员蛋白质组学:日本引领太空变革性研究。
IF 3.4 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-10 DOI: 10.1002/pmic.202300645
Alexia Tasoula, Nathaniel Szewczyk
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引用次数: 0
Editorial Board: Proteomics 20'24 编辑委员会:蛋白质组学 20'24
IF 3.4 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-10 DOI: 10.1002/pmic.202470162
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引用次数: 0
CoNglyPred: Accurate Prediction of N-Linked Glycosylation Sites Using ESM-2 and Structural Features With Graph Network and Co-Attention CoNglyPred:利用 ESM-2 和结构特征以及图形网络和共注意力准确预测 N-连接糖基化位点。
IF 3.4 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-03 DOI: 10.1002/pmic.202400210
Hongmei Wang, Long Zhao, Ziyuan Yu, Ximin Zeng, Shaoping Shi

N-Linked glycosylation is crucial for various biological processes such as protein folding, immune response, and cellular transport. Traditional experimental methods for determining N-linked glycosylation sites entail substantial time and labor investment, which has led to the development of computational approaches as a more efficient alternative. However, due to the limited availability of 3D structural data, existing prediction methods often struggle to fully utilize structural information and fall short in integrating sequence and structural information effectively. Motivated by the progress of protein pretrained language models (pLMs) and the breakthrough in protein structure prediction, we introduced a high-accuracy model called CoNglyPred. Having compared various pLMs, we opt for the large-scale pLM ESM-2 to extract sequence embeddings, thus mitigating certain limitations associated with manual feature extraction. Meanwhile, our approach employs a graph transformer network to process the 3D protein structures predicted by AlphaFold2. The final graph output and ESM-2 embedding are intricately integrated through a co-attention mechanism. Among a series of comprehensive experiments on the independent test dataset, CoNglyPred outperforms state-of-the-art models and demonstrates exceptional performance in case study. In addition, we are the first to report the uncertainty of N-linked glycosylation predictors using expected calibration error and expected uncertainty calibration error.

N-连接糖基化对蛋白质折叠、免疫反应和细胞运输等各种生物过程至关重要。确定N-连接糖基化位点的传统实验方法需要投入大量的时间和人力,因此人们开始开发计算方法作为更有效的替代方法。然而,由于三维结构数据有限,现有的预测方法往往难以充分利用结构信息,无法有效整合序列和结构信息。在蛋白质预训练语言模型(pLMs)取得进展和蛋白质结构预测取得突破的推动下,我们引入了一种名为 CoNglyPred 的高精度模型。在比较了各种 pLM 后,我们选择了大规模 pLM ESM-2 来提取序列嵌入,从而减少了人工特征提取的某些局限性。同时,我们的方法采用图转换器网络来处理 AlphaFold2 预测的三维蛋白质结构。最终的图输出和 ESM-2 嵌入通过共同关注机制错综复杂地结合在一起。在对独立测试数据集进行的一系列综合实验中,CoNglyPred 的表现优于最先进的模型,并在案例研究中表现出卓越的性能。此外,我们还首次使用预期校准误差和预期不确定性校准误差报告了N-连接糖基化预测因子的不确定性。
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引用次数: 0
Editorial Board: Proteomics 19'24 编辑委员会:蛋白质组学 19'24
IF 3.4 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-02 DOI: 10.1002/pmic.202470152
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引用次数: 0
Standard abbreviations 标准缩写。
IF 3.4 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-02 DOI: 10.1002/pmic.202470154
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引用次数: 0
Contents: Proteomics 19'24 内容:蛋白质组学 19'24
IF 3.4 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-02 DOI: 10.1002/pmic.202470153
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引用次数: 0
Identification of Key Genes in Fetal Gut Development at Single-Cell Level by Exploiting Machine Learning Techniques 利用机器学习技术在单细胞水平鉴定胎儿肠道发育过程中的关键基因
IF 3.4 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-26 DOI: 10.1002/pmic.202400104
QingLan Ma, Mei Meng, XianChao Zhou, Wei Guo, KaiYan Feng, Tao Huang, Yu-Dong Cai

The study of fetal gut development is critical due to its substantial influence on immediate neonatal and long-term adult health. Current research largely focuses on microbiome colonization, gut immunity, and barrier function, alongside the impact of external factors on these phenomena. Limited research has been dedicated to the categorization of developing fetal gut cells. Our study aimed to enhance our understanding of fetal gut development by employing advanced machine-learning techniques on single-cell sequencing data. This dataset consisted of 62,849 samples, each characterized by 33,694 distinct gene features. Four feature ranking algorithms were utilized to sort features according to their significance, resulting in four feature lists. Then, these lists were fed into an incremental feature selection method to extract essential genes, classification rules, and build efficient classifiers. Several important genes were recognized by multiple feature ranking algorithms, such as FGG, MDK, RBP1, RBP2, IGFBP7, and SPON2. These features were key in differentiating specific developing intestinal cells, including epithelial, immune, mesenchymal, and vasculature cells of the colon, duo jejunum, and ileum cells. The classification rules showed special gene expression patterns on some intestinal cell types and the efficient classifiers can be useful tools for identifying intestinal cells.

胎儿肠道发育对新生儿的近期健康和成年后的长期健康有着重大影响,因此对胎儿肠道发育的研究至关重要。目前的研究主要集中在微生物组定植、肠道免疫和屏障功能,以及外部因素对这些现象的影响。对发育中的胎儿肠道细胞进行分类的研究十分有限。我们的研究旨在通过对单细胞测序数据采用先进的机器学习技术,加深我们对胎儿肠道发育的了解。该数据集包括 62,849 个样本,每个样本都有 33,694 个不同的基因特征。研究人员利用四种特征排序算法根据特征的重要性对其进行排序,最终得出四个特征列表。然后,将这些列表输入增量特征选择方法,以提取重要基因和分类规则,并建立高效的分类器。多个特征排序算法识别出了几个重要基因,如 FGG、MDK、RBP1、RBP2、IGFBP7 和 SPON2。这些特征是区分特定发育中肠细胞的关键,包括结肠、空肠和回肠细胞的上皮细胞、免疫细胞、间质细胞和血管细胞。分类规则显示了某些肠细胞类型的特殊基因表达模式,高效的分类器可作为识别肠细胞的有用工具。
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引用次数: 0
Development of a Proteomic Workflow for the Identification of Heparan Sulphate Proteoglycan-Binding Substrates of ADAM17 开发用于鉴定 ADAM17 的硫酸肝素蛋白多糖结合底物的蛋白质组工作流程。
IF 3.4 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-24 DOI: 10.1002/pmic.202400076
Matteo Calligaris, Donatella Pia Spanò, Maria Chiara Puccio, Stephan A. Müller, Simone Bonelli, Margot Lo Pinto, Giovanni Zito, Carl P. Blobel, Stefan F. Lichtenthaler, Linda Troeberg, Simone Dario Scilabra

Ectodomain shedding, which is the proteolytic release of transmembrane proteins from the cell surface, is crucial for cell-to-cell communication and other biological processes. The metalloproteinase ADAM17 mediates ectodomain shedding of over 50 transmembrane proteins ranging from cytokines and growth factors, such as TNF and EGFR ligands, to signalling receptors and adhesion molecules. Yet, the ADAM17 sheddome is only partly defined and biological functions of the protease have not been fully characterized. Some ADAM17 substrates (e.g., HB-EGF) are known to bind to heparan sulphate proteoglycans (HSPG), and we hypothesised that such substrates would be under-represented in traditional secretome analyses, due to their binding to cell surface or pericellular HSPGs. Thus, to identify novel HSPG-binding ADAM17 substrates, we developed a proteomic workflow that involves addition of heparin to solubilize HSPG-binding proteins from the cell layer, thereby allowing their mass spectrometry detection by heparin-treated secretome (HEP-SEC) analysis. Applying this methodology to murine embryonic fibroblasts stimulated with an ADAM17 activator enabled us to identify 47 transmembrane proteins that were shed in response to ADAM17 activation. This included known HSPG-binding ADAM17 substrates (i.e., HB-EGF, CX3CL1) and 14 novel HSPG-binding putative ADAM17 substrates. Two of these, MHC-I and IL1RL1, were validated as ADAM17 substrates by immunoblotting.

外膜脱落是跨膜蛋白从细胞表面的蛋白水解释放,对于细胞间通信和其他生物过程至关重要。金属蛋白酶 ADAM17 可介导 50 多种跨膜蛋白的外膜脱落,包括细胞因子和生长因子(如 TNF 和表皮生长因子受体配体)、信号受体和粘附分子。然而,ADAM17 的脱落组仅得到部分界定,该蛋白酶的生物功能也尚未完全确定。已知一些 ADAM17 底物(如 HB-EGF)会与硫酸肝素蛋白多糖(HSPG)结合,我们推测这类底物由于会与细胞表面或细胞周围的 HSPGs 结合,因此在传统的分泌物组分析中代表性不足。因此,为了鉴定新型HSPG结合ADAM17底物,我们开发了一种蛋白质组学工作流程,其中包括添加肝素以溶解细胞层中的HSPG结合蛋白,从而通过肝素处理分泌物组(HEP-SEC)分析对其进行质谱检测。将这种方法应用于受到 ADAM17 激活剂刺激的小鼠胚胎成纤维细胞,使我们能够鉴定出 47 种因 ADAM17 激活而脱落的跨膜蛋白。其中包括已知的与 HSPG 结合的 ADAM17 底物(即 HB-EGF、CX3CL1)和 14 种新型的与 HSPG 结合的推测 ADAM17 底物。其中两个底物(MHC-I 和 IL1RL1)通过免疫印迹验证为 ADAM17 底物。
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引用次数: 0
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Proteomics
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