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1. chinaXiv:202002.00082 [pdf]

ACE2 shedding and furin abundance in target organs may influence the efficiency of SARS-CoV-2 entry

Yuanchen Ma; Yinong Huang; Tao Wang; Andy Peng Xiang; Weijun Huang
Subjects: Medicine, Pharmacy >> Preclinical Medicine

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a recently identified lineage B coronavirus, causing rapid worldwide outbreak of Corona Virus Disease 2019 (COVID-19). Despite genetically closed to SARS-CoV, SARS-CoV-2 seems to possess enhanced infectivity and subtle different clinical features, which may hamper the early screening of suspected patients as well as the control of virus transmission. Unfortunately, there are few tools to predict the potential target organ damage and possible clinical manifestations caused by such novel coronavirus. To solve this problem, we investigate the potential host cell entry mechanisms of SARS-CoV-2 through bioinformatics. Using the online single-cell sequence datasets, we analyze the expression of major receptor in host cells that mediates the virus entry, including angiotensin converting enzyme 2 (ACE2), and its co-expressed membrane endopeptidases. The results indicated the differential expression of ADAM10 and ADAM17 might contribute to the ACE2 shedding and affect the membrane ACE2 abundance. We further confirm a putative furin-cleavage site reported recently in the spike protein of SARS-CoV-2, which may facilitate the virus-cell fusion. Based on these findings, we develop a novel approach that comprehensively analyzed the virus receptor expression, ACE2 shedding, membrane fusion activity, virus uptake and virus replication to evaluate the infectivity of SARS-CoV-2 to different human organs. Our results indicate that, in addition to airway epithelia, cardiac tissue and enteric canals are susceptible to SARS-CoV-2 as well.

submitted time 2020-02-27 Hits44632Downloads4666 Comment 0

2. chinaXiv:202002.00062 [pdf]

Furin, a potential therapeutic target for COVID-19

Hua Li; Canrong Wu; Yueying Yang; Yang Liu; Peng Zhang; Yali Wang; Qiqi Wang; Yang Xu; Mingxue Li; Mengzhu Zheng; Lixia Chen
Subjects: Medicine, Pharmacy >> Pharmacology

A novel coronavirus (SARS-CoV-2) infectious disease has broken out in Wuhan, Hubei Province since December 2019, and spread rapidly from Wuhan to other areas, which has been listed as an international concerning public health emergency. We compared the Spike proteins from four sources, SARS-CoV-2, SARS-CoV, MERS-CoV and Bat-CoVRaTG13, and found that the SARS-CoV-2 virus sequence had redundant PRRA sequences. Through a series of analyses, we propose the reason why SARS-CoV-2is more infectious than other coronaviruses. And through structure based virtual ligand screening, we foundpotentialfurin inhibitors, which might be used in the treatment of new coronary pneumonia.

submitted time 2020-02-23 Hits137726Downloads19280 Comment 0

3. chinaXiv:202002.00004 [pdf]

武汉2019冠状病毒S蛋白可能存在Furin蛋白酶切位点

李鑫; 段广有; 张伟; 施劲松; 陈嘉源; 陈舜梅; 高山; 阮吉寿
Subjects: Biology >> Virology

摘要:2019年12月,中国武汉报道了2019新型冠状病毒(2019 novel Coronavirus,2019-nCoV)引起的肺炎。基于基因组信息,我们前期研究结果显示2019-nCoV与SARS冠状病毒虽然同属于Beta冠状病毒B亚群(BB冠状病毒),但两种病毒差异很大,这一结果与两者临床症状差异一致。前期研究还发现了BB冠状病毒存在大量的可变翻译,并从分子水平揭示了BB冠状病毒变异快、多样性高的特点。本研究在国际上首次报道BB冠状病毒S蛋白上的一个重要突变,这个突变使2019-nCoV具有了一个可供Furin蛋白酶切的位点,是除鼠肝炎冠状病毒外所有的其它BB冠状病毒(包括SARS和SARS样(SARS-like)冠状病毒)所不具有的。这个突变有可能增强了2019-nCoV侵染细胞的效率,进而使其传播力显著大于SARS冠状病毒。由于这个突变,2019冠状病毒的包装机制也会不同于SARS等其它大部分Beta冠状病毒,而有可能与鼠肝炎冠状病毒、HIV、埃博拉病毒和一些禽流感病毒的包装机制相同。作为一个意外发现,一些禽流感病毒也可以通过突变获得Furin蛋白酶切位点。对这个重要突变的后续研究将为揭示2019-nCoV传播力强的原因,以及为药物、抗体和疫苗的开发等工作奠定基础。

submitted time 2020-02-14 Hits90068Downloads17274 Comment 0

4. chinaXiv:201712.01121 [pdf]

Furin抑制剂α1-PDX对宫颈癌HeLa细胞生长、转移和成瘤的影响

石冲(1);张国彬(2);韩宝生(1);杨俊红(1);刘恒(1);习瑾昆(2)
Subjects: Medicine, Pharmacy >> Preclinical Medicine

目的研究α1-PDX对宫颈癌细胞株HeLa细胞生长、转移及成瘤的影响并探讨其机制。方法α1-PDX转染HeLa细胞,通过MTT细胞增殖实验,Boyden细胞迁移和侵袭实验观察HeLa细胞的生长,迁移和侵袭能力。采用蛋白印迹法检测细胞Furin及产物膜性I型金属蛋白酶(MT1-MMP)蛋白量的变化,荧光酶底物实验检测Furin活性。制作HeLa细胞致瘤裸鼠模型研究肿瘤体内生长。结果细胞增殖实验结果显示,与对照组相比24 h α1-PDX组HeLa细胞体外生长抑制18.4%,差异有统计学意义(P<0.01);细胞迁移和侵袭实验结果显示,α1-PDX处理组穿过滤膜和基质胶的HeLa细胞数量均明显少于对照组(P<0.01) ;HeLa/PDX可显著降低Furin的活性和MTI-MMP的蛋白水平。HeLa/PDX致瘤裸鼠的成瘤机率及皮下肿瘤体积明显小于对照组。结论α1-PDX可有效抑制人宫颈癌HeLa细胞的生长、转移及成瘤能力,机制可能与降低Furin活性及产物MTI-MMP的表达有关。

submitted time 2017-12-07 From cooperative journals:《南方医科大学学报》 Hits694Downloads422 Comment 0

5. chinaXiv:201705.00713 [pdf]

Spike Protein, S, of Human Coronavirus HKU1: Role in Viral Life Cycle and Application in Antibody Detection

Chan, Che-Man; Woo, Patrick C. Y; Lau, Susanna K. P; Tse, Herman; Chen, Hong-Lin; Li, Feng; Zheng, Bo-Jian; Chen, Ling; Huang, Jian-Dong; Yuen, Kwok-Yung
Subjects: Biology >> Biomedical Laboratory Science

We recently described the discovery, genome, clinical features, genotypes and evolution of a novel and global human respiratory virus named human coronavirus HKU1 (HCoV-HKU1) which is not yet culturable. We expressed a C-terminal FLAG-tagged CoV-HKU1 spike (S) protein by the Semliki Forest Virus (SFV) system and investigated its maturation profile. Pulse chase labeling revealed that S-FLAG was expressed as high-mannose N-glycans of monomers and trimers. It was predominantly cleaved into subdomains S1 and S2 during maturation. S1 was secreted into the medium. Immunofluorescence analysis visualized S along the secretory pathway from endoplasmic reticulum to plasma membrane. Cleavage of S and release of HCoV-HKU1 S pseudotyped virus were inhibited by furin or furin-like enzyme inhibitors. The cell-based expressed full-length S-FLAG could be recognized by the convalescent serum obtained from a patient with HCoV-HKU1 pneumonia. The data suggest that the native form of HCoV-HKU1 spike expressed in our system can be used in developing serological diagnostic assay and in understanding the role of S in the viral life cycle. Exp Biol Med 233:1527-1536, 2008

submitted time 2017-05-10 Hits1233Downloads657 Comment 0

6. chinaXiv:201605.00465 [pdf]

Co‐expression of the protease furin in Nicotiana benthamiana leads to efficient processing of latent transforming growth factor‐β1 into a biologically active protein

Ruud H. P. Wilbers; Lotte B. Westerhof; Debbie R. van Raaij; Marloes van Adrichem; Andreas D. Prakasa; Jose L. Lozano-Torres; Jaap Bakker; Geert Smant; Arjen Schots
Subjects: Biology >> Botany >> Plant biochemistry, plant Biophysics

Transforming growth factor beta (TGF-β) is a signalling molecule that plays a key role in developmental and immunological processes in mammals. Three TGF-β isoforms exist in humans, and each isoform has unique therapeutic potential. Plants offer a platform for the production of recombinant proteins, which is cheap and easy to scale up and has a low risk of contamination with human pathogens. TGF-β3 has been produced in plants before using a chloroplast expression system. However, this strategy requires chemical refolding to obtain a biologically active protein. In this study, we investigated the possibility to transiently express active human TGF-β1 in Nicotiana benthamiana plants. We successfully expressed mature TGF-β1 in the absence of the latency-associated peptide (LAP) using different strategies, but the obtained proteins were inactive. Upon expression of LAP-TGF-β1, we were able to show that processing of the latent complex by a furin-like protease does not occur in planta. The use of a chitinase signal peptide enhanced the expression and secretion of LAP-TGF-β1, and co-expression of human furin enabled the proteolytic processing of latent TGF-β1. Engineering the plant post-translational machinery by co-expressing human furin also enhanced the accumulation of biologically active TGF-β1. This engineering step is quite remarkable, as furin requires multiple processing steps and correct localization within the secretory pathway to become active. Our data demonstrate that plants can be a suitable platform for the production of complex proteins that rely on specific proteolytic processing.

submitted time 2016-05-04 Hits1706Downloads982 Comment 0

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