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9. Progress of liquid chromatography and mass spectrometry-based single-cell proteomics

Author

Huo Zian, Guo Yueshuai, Wang Yue, Situ Chenghao, and Guo Xuejiang

Subjects
single cell, proteomics, mass spectrometry (ms), liquid chromatography (lc), lc-ms, Medicine, Biotechnology, TP248.13-248.65
Abstract

Proteins are the main functional executor of cellular function. Due to the fact that proteins could not be amplified in vitro, the development of single-cell proteomics technology lags behind single-cell genomics and transcriptomics technologies. The traditional proteomics technology detects the average protein expression levels from large amounts of cells; nevertheless, it is unable to address the cell subtypes and heterogeneity among individual cells. Analysis of proteins at the single-cell level can help characterize the molecular basis of cell phenotypes and heterogeneity. With the rapid development of mass spectrometers, mass spectrometry-based methods have pushed single-cell proteomics to a new level. In this review, we summarized the recent research progress of single-cell proteomics methods based on LC-MS/MS, including single cell selection, sample preparation, isotope labelling technology, peptide separation, mass spectrometry analysis and data analysis, and its applications in biomedical research. The challenges and prospects of single-cell proteomics in the future are also prospected. Advances in single cell proteomics will provide new ideas and methods for biomedical research and deepen our understanding of human health and disease.

Published
2022
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19. The Formation of Bixbyite-Type Mn2O3 via Pyocyanin-Dependent Mn(II) Oxidation of Soil-Derived Pseudomonas aeruginosa.

Author

Xue, Lan, Liao, Fengfeng, Tang, Fengzhen, Chen, Sha, Hu, Mingzhen, Qiao, Liying, Guo, Yueshuai, Sun, Huatao, and Li, Ding

Subjects
CULTURE media (Biology), OXIDATION, CELL culture, PSEUDOMONAS, CELL suspensions, REACTIVE oxygen species, PSEUDOMONAS aeruginosa
Abstract

Bacterial Mn(II) oxidation is believed to play a dominant role in accelerating the rate of Mn biomineralization in nature. Commonly, bacteria employ two ways involving Mn(II) oxidases and reactive oxygen species to oxidize Mn(II). In this study, a new strategy for bacterial Mn(II) oxidation involving the pyocyanin, a greenish blue phenazine pigment from Pseudomonas aeruginosa, was discovered. To begin with, a bacterial strain L3 was isolated from soils and identified as Pseudomonas aeruginosa, which exhibited the ability of Mn(II) oxidation. Next, the pyocyanin was purified from strain L3 cultures and proven to be involved in Mn(II) oxidation. Particularly, the oxidation of Mn(II) by pyocyanin was dependent on its ambient pH. In comparison with pH of 5 and 7, pyocyanin (the initial value of OD387 was 0.56 at pH of 2) showed a stronger capability of oxidizing Mn(II) at pH of 9, reaching 144.03 μg L−1 of Mn oxides after 108 h of Mn(II) oxidation, while pyocyanin ultimately produced 43.81 μg L−1 at pH of 7 and 3.32 μg L−1 at pH of 5, respectively. Further, strain L3 cultures were fractionated into three parts, i.e., the cell culture solution, fermentation supernatant, and cell suspension, and the Mn(II)-oxidizing activity was found to be distributed in the cell culture solution and fermentation supernatant, as evidenced by the formation of blackish glossy Mn oxides. Specifically, in the first half, the rate of Mn(II) oxidation by the fermentation supernatant was higher than that by the cell culture solution, whereas in the second half, the cell culture solution showed the much higher Mn(II)-oxidizing activity compared to the fermentation supernatant. Last but not least, the collective results from mineral characterization demonstrated that, the Mn oxides produced by P. aeruginosa strain L3, either by the cell culture solution or by the fermentation supernatant, were bixbyite-type Mn2O3 with poor crystallinity. [ABSTRACT FROM AUTHOR]

Published
2024
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29. Additional file 2 of Regulation of Miwi-mediated mRNA stabilization by Ck137956/Tssa is essential for male fertility

Author

Chen, Yu, Zhang, Xiangzheng, Jiang, Jiayin, Luo, Mengjiao, Tu, Haixia, Xu, Chen, Tan, Huanhuan, Zhou, Xin, Chen, Hong, Han, Xudong, Yue, Qiuling, Guo, Yueshuai, Zheng, Ke, Qi, Yaling, Situ, Chenghao, Cui, Yiqiang, and Guo, Xuejiang

Abstract

Additional file 2: Table S1. siRNA oligonucleotides. Table S2. Gene-specific primers used in qRT-PCR. Table S3. piRNA specific primers used in qRT-PCR. Table S4. Antibodies used in this study.

Published
2023
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31. Homeodomain-interacting protein kinase HIPK4 regulates phosphorylation of manchette protein RIMBP3 during spermiogenesis

Author

Liu, Xiaofei, primary, Zang, Chunyan, additional, Wu, Yifei, additional, Meng, Ru, additional, Chen, Yu, additional, Jiang, Tao, additional, Wang, Cheng, additional, Yang, Xiaoyu, additional, Guo, Yueshuai, additional, Situ, Chenghao, additional, Hu, Zhibin, additional, Zhang, Jun, additional, and Guo, Xuejiang, additional

Published
2022
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34. SARS-CoV-2 ORF10 impairs cilia by enhancing CUL2ZYG11B activity

Author

Wang, Liying, primary, Liu, Chao, additional, Yang, Bo, additional, Zhang, Haotian, additional, Jiao, Jian, additional, Zhang, Ruidan, additional, Liu, Shujun, additional, Xiao, Sai, additional, Chen, Yinghong, additional, Liu, Bo, additional, Ma, Yanjie, additional, Duan, Xuefeng, additional, Guo, Yueshuai, additional, Guo, Mengmeng, additional, Wu, Bingbing, additional, Wang, Xiangdong, additional, Huang, Xingxu, additional, Yang, Haitao, additional, Gui, Yaoting, additional, Fang, Min, additional, Zhang, Luo, additional, Duo, Shuguang, additional, Guo, Xuejiang, additional, and Li, Wei, additional

Published
2022
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37. SARS-CoV-2 ORF10 Impairs Cilia by Enhancing CUL2ZYG11B Activity

Author

Wang, Liying, Liu, Chao, Yang, Bo, Zhang, Haotian, Jiao, Jiao, Zhang, Ruidan, Liu, Shujun, Xiao, Sai, Chen, Yinghong, Liu, Bo, Ma, Yanjie, Duan, Xuefeng, Guo, Yueshuai, Guo, Mengmeng, Wu, Bingbing, Wang, Xiangdong, Huang, Xingxu, Yang, Haitao, Gui, Yaoting, Fang, Min, Zhang, Luo, Duo, Shuguang, Guo, Xuejiang, Li, Wei, Wang, Liying, Liu, Chao, Yang, Bo, Zhang, Haotian, Jiao, Jiao, Zhang, Ruidan, Liu, Shujun, Xiao, Sai, Chen, Yinghong, Liu, Bo, Ma, Yanjie, Duan, Xuefeng, Guo, Yueshuai, Guo, Mengmeng, Wu, Bingbing, Wang, Xiangdong, Huang, Xingxu, Yang, Haitao, Gui, Yaoting, Fang, Min, Zhang, Luo, Duo, Shuguang, Guo, Xuejiang, and Li, Wei

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causal pathogen of the ongoing global pandemic of coronavirus disease 2019 (COVID-19). Loss of smell and taste are symptoms of COVID-19, and may be related to cilia dysfunction. Here, we found that the SARS-CoV-2 ORF10 increases the overall E3 ligase activity of the CUL2ZYG11B complex by interacting with ZYG11B. Enhanced CUL2ZYG11B activity by ORF10 causes increased ubiquitination and subsequent proteasome-mediated degradation of an intraflagellar transport (IFT) complex B protein, IFT46, thereby impairing both cilia biogenesis and maintenance. Further, we show that exposure of the respiratory tract of hACE2 mice to SARS-CoV-2 or SARS-CoV-2 ORF10 alone results in cilia-dysfunction-related phenotypes, and the ORF10 expression in primary human nasal epithelial cells (HNECs) also caused a rapid loss of the ciliary layer. Our study demonstrates how SARS-CoV-2 ORF10 hijacks CUL2ZYG11B to eliminate IFT46 and leads to cilia dysfunction, thereby offering a powerful etiopathological explanation for how SARS-CoV-2 causes multiple cilia-dysfunction-related symptoms specific to COVID-19. © 2022 Wang et al.

Published
2022

40. Integrative proteome analysis implicates aberrant RNA splicing in impaired developmental potential of aged mouse oocytes

Author

Li, Mingrui, primary, Ren, Chao, additional, Zhou, Shuai, additional, He, Yuanlin, additional, Guo, Yueshuai, additional, Zhang, Hao, additional, Liu, Lu, additional, Cao, Qiqi, additional, Wang, Congjing, additional, Huang, Jie, additional, Hu, Yue, additional, Bai, Xue, additional, Guo, Xuejiang, additional, Shu, Wenjie, additional, and Huo, Ran, additional

Published
2021
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42. Maternal RNF114-mediated target substrate degradation regulates zygotic genome activation in mouse embryos

Author

Zhou, Shuai, primary, Guo, Yueshuai, additional, Sun, Haifeng, additional, Liu, Lu, additional, Yao, Liping, additional, Liu, Chao, additional, He, Yuanlin, additional, Cao, Shanren, additional, Zhou, Cheng, additional, Li, Mingrui, additional, Cao, Yumeng, additional, Wang, Congjing, additional, Lu, Qianneng, additional, Li, Wei, additional, Guo, Xuejiang, additional, and Huo, Ran, additional

Published
2021
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44. Quantitative proteomic biomarkers from extracellular vesicles of human seminal plasma in the differential diagnosis of azoospermia

Author

Yao, Liping, primary, Guo, Yueshuai, additional, Zhang, Xiangzheng, additional, Xu, Chen, additional, Wang, Yichun, additional, Liu, Xiaofei, additional, Wang, Yue, additional, Li, Yan, additional, Qi, Yaling, additional, Sha, Jiahao, additional, Qin, Chao, additional, Yang, Xiaoyu, additional, and Guo, Xuejiang, additional

Published
2021
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45. Characterization of Metabolic Patterns in Mouse Oocytes during Meiotic Maturation

Author

Li, Ling, primary, Zhu, Shuai, additional, Shu, Wenjie, additional, Guo, Yueshuai, additional, Guan, Yusheng, additional, Zeng, Juan, additional, Wang, Haichao, additional, Han, Longsen, additional, Zhang, Jiaqi, additional, Liu, Xiaohui, additional, Li, Chunling, additional, Hou, Xiaojing, additional, Gao, Min, additional, Ge, Juan, additional, Ren, Chao, additional, Zhang, Hao, additional, Schedl, Tim, additional, Guo, Xuejiang, additional, Chen, Minjian, additional, and Wang, Qiang, additional

Published
2020
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48. m6A reader protein YTHDF2 regulates spermatogenesis by timely clearance of phase‐specific transcripts.

Author

Qi, Meijie, Sun, Haifeng, Guo, Yueshuai, Zhou, Yu, Gu, Xueying, Jin, Jiachuan, Chen, Xiaoxu, Wang, Fangzhu, Ma, Honghui, Guo, Xuejiang, Chen, Hao, and Shen, Bin

Subjects
GERM cells, KNOCKOUT mice, SPERMATOGENESIS, WESTERN immunoblotting, SPERM competition, SPERMATOZOA, LABORATORY mice
Abstract

Objectives: Accumulating evidences show that the regulatory network of m6A modification is essential for mammalian spermatogenesis. However, as an m6A reader, the roles of YTHDF2 remain enigmatic due to the lack of a proper model. Here, we employed the germ cell conditional knockout mouse model and explored the function of YTHDF2 in spermatogenesis. Materials and methods: Ythdf2 germ cell conditional knockout mice were obtained by crossing Ythdf2‐floxed mice with Vasa‐Cre and Stra8‐Cre mice. Haematoxylin and eosin (HE) staining, immunofluorescent staining and Western blotting were used for phenotyping. CASA, IVF and ICSI were applied for sperm function analysis. RNA‐seq, YTHDF2‐RIP‐seq and quantitative real‐time PCR were used to explore transcriptome changes and molecular mechanism analysis. Results: Our results showed that YTHDF2 was highly expressed in spermatogenic cells. The germ cell conditional knockout males were sterile, and their sperm displayed malformation, impaired motility, and lost fertilization ability. During differentiated spermatogonia transiting to pachytene spermatocyte, most m6A‐modified YTHDF2 targets that were degraded in control germ cells persisted in pachytene spermatocytes of Ythdf2‐vKO mice. These delayed mRNAs were mainly enriched in pathways related to the regulation of transcription, and disturbed the transcriptome of round spermatid and elongated spermatid subsequently. Conclusion: Our data demonstrate that YTHDF2 facilitates the timely turnover of phase‐specific transcripts to ensure the proper progression of spermatogenesis, which highlights a critical role of YTHDF2 in spermatogenesis. [ABSTRACT FROM AUTHOR]

Published
2022
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49. Proteomic Analysis of Dpy19l2‐Deficient Human Globozoospermia Reveals Multiple Molecular Defects

Author

Guo, Yueshuai, primary, Jiang, Jiayin, additional, Zhang, Haotian, additional, Wen, Yang, additional, Zhang, Hao, additional, Cui, Yiqiang, additional, Tian, Jianyu, additional, Jiang, Min, additional, Liu, Xiaofei, additional, Wang, Gaigai, additional, Li, Yan, additional, Hu, Zhibin, additional, Zhou, Zuomin, additional, Sha, Jiahao, additional, Chen, Daozhen, additional, Yang, Xiaoyu, additional, and Guo, Xuejiang, additional

Published
2019
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50. Myotubularin related protein 7 is essential for the spermatogonial stem cell homeostasis via PI3K/AKT signaling

Author

Zhao, Dan, primary, Shen, Cong, additional, Gao, Tingting, additional, Li, Hong, additional, Guo, Yueshuai, additional, Li, Feng, additional, Liu, Chenchen, additional, Liu, Yuanyuan, additional, Chen, Xia, additional, Zhang, Xi, additional, Wu, Yangyang, additional, Yu, Yi, additional, Lin, Meng, additional, Yuan, Yan, additional, Chen, Xiaofang, additional, Huang, Xiaoyan, additional, Yang, Shenmin, additional, Yu, Jun, additional, Zhang, Jun, additional, and Zheng, Bo, additional

Published
2019
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