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High-coverage proteome analysis reveals the first insight of protein modification systems in the pathogenic spirochete Leptospira interrogans 被引量:8
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作者 Xing-Jun Cao Jie Dai +10 位作者 Hao Xu Song Nie Xiao Chang Bao-Yu Hu Quan-Hu Sheng Lian-Shui Wang Zhi-Bin Ning Yi-Xue Li Xiao-Kui Guo Guo-Ping Zhao Rong Zeng 《Cell Research》 SCIE CAS CSCD 2010年第2期197-210,共14页
Leptospirosis is a widespread zoonotic disease caused by pathogenic spirochetes of the genus Leptospira that infects humans and a wide range of animals. By combining computational prediction and high-accuracy tandem m... Leptospirosis is a widespread zoonotic disease caused by pathogenic spirochetes of the genus Leptospira that infects humans and a wide range of animals. By combining computational prediction and high-accuracy tandem mass spectra, we revised the genome annotation of Leptospira interrogans serovar Lai, a free-living pathogenic spirochete responsible for leptospirosis, providing substantial peptide evidence for novel genes and new gene boundaries. Subsequently, we presented a high-coverage proteome analysis of protein expression and multiple posttranslational modifications (PTMs). Approximately 64.3% of the predicted L. interrogans proteins were cataloged by detecting 2 540 proteins. Meanwhile, a profile of multiple PTMs was concurrently established, containing in total 32 phosphorylated, 46 acetylated and 155 methylated proteins. The PTM systems in the serovar Lai show unique features. Unique eukaryotic-like features of L. interrogans protein modifications were demonstrated in both phosphorylation and arginine methylation. This systematic analysis provides not only comprehensive information of high-coverage protein expression and multiple modifications in prokaryotes but also a view suggesting that the evolutionarily primitive L. interrogans shares significant similarities in protein modification systems with eukaryotes. 展开更多
关键词 leptospira interrogans posttranslational modification eukaryotic-like evolutionary conservation
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Severe community-acquired pneumonia caused by Leptospira interrogans:A case report and review of literature 被引量:2
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作者 Qiu-Hong Bao Li Yu +4 位作者 Jian-Jun Ding Ying-Jun Chen Jun-Wei Wang Jian-Ming Pang Qi Jin 《World Journal of Clinical Cases》 SCIE 2021年第8期1901-1908,共8页
BACKGROUND Leptospira is an uncommon pathogen for adult severe community-acquired pneumonia and its nonspecific manifestations and limited diagnostic tests make it difficult to identify.Although conventional penicilli... BACKGROUND Leptospira is an uncommon pathogen for adult severe community-acquired pneumonia and its nonspecific manifestations and limited diagnostic tests make it difficult to identify.Although conventional penicillin remains efficacious to treat leptospirosis,failure in early diagnosis and treatment can lead to progression into a deadly syndrome with multiple organ dysfunction.Next generation sequencing is of great value to understand cases with infection of unknown cause,which could help in the diagnosis of uncertain Leptospira infection.CASE SUMMARY We recently managed a patient with fever,cough and dyspnea on admission that progressed into persistent adult respiratory distress syndrome,hemoptysis and hematuria after admission.In this case,the rare Leptospira infection was clouded by the positive influenza tests at admission,delaying early Leptospira-targeted antibiotics administration.Next generation sequencing,a novel molecular diagnostic tool,provided a key hint to uncover the crucial pathogen,Leptospira interrogans,further supported by the possible occupational exposure history.Subsequent conventional penicillin and mechanical respiratory support were administrated to cure the patient successfully without any sequela.CONCLUSION Clinicians must pay attention to possible exposure history and keep uncommon Leptospira in mind when managing pneumonia with unknown causes. 展开更多
关键词 leptospira interrogans PNEUMONIA Next generation sequencing INFLUENZA DIAGNOSIS Case report
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THE CONSTRUCTION AND EXPRESSION OF RECOMBINANT SHUTTLE PLASMID WITH OMPL1 GENE FROM LEPTOSPIRA INTERROGANS SEROVAR LAI STRAIN 017 IN BACILLE CALMETTE-GUERIN 被引量:2
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作者 鲍朗 邱洪宇 +2 位作者 晏菊芳 谢勇恩 陈玮 《Chinese Medical Sciences Journal》 CAS CSCD 2002年第2期81-84,共4页
Objective.To construct recombinant BCG again st leptospirosis.Methods.We amplified the entire open readin g frame of the OmpL1gene from the genome of the leptospire serovar Lai strain 017.Two recombin ant plasmids pBQ... Objective.To construct recombinant BCG again st leptospirosis.Methods.We amplified the entire open readin g frame of the OmpL1gene from the genome of the leptospire serovar Lai strain 017.Two recombin ant plasmids pBQ1and pBQ2were constructed by oriented ligation based on the E.coli-BCG shuttle plasmids pMV261and pMV361respectively.The recombinant plasmids were transformed into BCG by electroporation.The rBCGs bearing pBQ1and pBQ2were induced by high temperature of 45℃.Results.The expressed product,a 35kD prote in was detected by SDS-PAGE.The resu lt indicates that pBQ1and pBQ2can express OmpL1in rBCG.Conclusion.The technical methods in this study may help detect the immunogenicity a nd immunoprotection of OmpL1and develop more safe,highl y effective rBCG bearing leptospira l antigen with long-lasting protection. 展开更多
关键词 leptospira interrogans serovar Lai recombinant BCG OmpL1gene
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Production of reactive oxygen species and expression of inducible nitric oxide synthase in rat isolated Kupffer cells stimulated by Leptospira interrogans and Borrelia burgdorfen 被引量:1
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作者 Antonella Marangoni Silvia Accardo +6 位作者 Rita Aldini Massimo Guardigli Francesca Cavrini Vittorio Sambri Marco Montagnani Aldo Roda Roberto Cevenini 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第19期3077-3081,共5页
AIM: To evaluate the production of reactive oxygen species (ROS) and the expression of inducible nitric oxide synthase (iNOS) in rat isolated Kupffer cells (KCs) stimulated by Leptospira interrogans and Borreli... AIM: To evaluate the production of reactive oxygen species (ROS) and the expression of inducible nitric oxide synthase (iNOS) in rat isolated Kupffer cells (KCs) stimulated by Leptospira interrogans and Borrelia burgdorferi. METHODS: Rat Kupffer cells were separated by perfusion of the liver with 0.05% collagenase, and purified by Percoll gradients. Pudfied Kupffer cells were tested in vitro with alive L.interogans and B. burgdorferi preparations. The production of ROS was determined by chemiluminescence, whereas iNOS protein expression was evaluated by Western blot assay using anti-iNOS antibodies. RESULTS: B. burgdorferi and to a less extent L. interrogans induced ROS production with a peak 35 min after infection. The chemiluminescence signal progressively diminished and was undetectable by 180 min of incubation. Leptospirae and borreliae induced an increased iNOS expression in Kupffer cells that peaked at 6 hours and was still evident 22 h after infection. CONCLUSION: Both genera of spirochetes induced ROS and iNOS production in rat Kupffer cells. Since the cause of liver damage both in leptospiral as well as in borrelial infections are still unknown, we suggest that leptospira and borrelia damage of the liver can be initially mediated by oxygen radicals, and is then maintained at least in part by nitric oxide. 展开更多
关键词 leptospira interrogans Borrelia burgdorferi Inducible nitric oxide synthase Nitric oxide Reactive oxygen species Kupffer cells CHEMILUMINESCENCE
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Molecular cloning of genes flhA and flhB_2 for flagellar biosynthesis of Leptospira interrogans and functional prediction of the prokaryotic expressing products
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作者 XIN YING WANG JIE YAN +1 位作者 DONG JIAO LUO HONG XUE FAN 《Journal of Microbiology and Immunology》 2005年第3期224-231,共8页
To determine the pathogenic potential of the leptospiral flagella-associated proteins, the genes flhA and flhB2 encoding the biosynthesis of flagella of leptospira interrogans serogroup icterohaemor- rhagiae serovar l... To determine the pathogenic potential of the leptospiral flagella-associated proteins, the genes flhA and flhB2 encoding the biosynthesis of flagella of leptospira interrogans serogroup icterohaemor- rhagiae serovar lai stain 56601 were cloned and their prokaryotic expression systems were constructed. It was demonstrated that the cloned flhA and flhB2 genes had 2118 bp in length and showed 100% and 99.9% of homologies in their nucleotide sequences and 100% and 98.8% of homologies in their putative amino acid sequences respectively, in comparison with those of previously reported. The prokaryotic expression systems under the induction with IPTG could efficiently express the target proteins rFlhA and rFlhB2 with the outputs of approximate 10% of the total bacterial proteins. Based on the sequences of the cloned genes flhA and flhB2, the structural features in associated with pathogenesis and the functions of the target proteins were analyzed with bioinformatics softwares, in which the FlhA was found to have 7 major transmembrane helices, while the FlhB2 had 5 ones. The conserved domains in the FlhA showed high similarity to those of the FHIPEP of the other bacterial FlhA and EscV families, but the conserved domains in the FlhB2 were similar to those of bac-export-2 and EscU families, EscV and EscU families being the protein products of the type IR secretion system in association with pathogenesis. The FlhA and FlhB2 also contained protein kinase C (PKC) and protein tyrosine kinase (PTK) phosphorylation sites, indicating that PKC and PTK of host cells were involved in the internalization and intracellular proliferation in the pathogenesis of microorganisms. All these data leads to a conclusion that the flhA and flhB2 genes of L. interrogans are relatively conserved and their gene products have great potential in the pathogenesis of this organism. 展开更多
关键词 leptospira interrogans flhA and flhB2 genes Clone/expression Pathogenesis/prediction
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A molecular dynamics simulation study of peptide deformylase from Leptospira interrogans complex:Exploring the closing mechanism of the substrate pocket
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作者 Qiang Wang Jian Wu Wang +1 位作者 Zheng Ting Cai Wei Ren Xu 《Chinese Chemical Letters》 SCIE CAS CSCD 2008年第4期497-500,共4页
To explore the closing mechanism of the substrate pocket,we perform a 16,000 ps molecular dynamics simulation separately on the ligand-free and actinonin-bound peptide deformylase from Leptospira interrogans.Our resul... To explore the closing mechanism of the substrate pocket,we perform a 16,000 ps molecular dynamics simulation separately on the ligand-free and actinonin-bound peptide deformylase from Leptospira interrogans.Our results show that the CD-loop, hydrophilic inhibitor and hydrophobic cluster are necessary for the formation of semi-open conformation,and Tyr71 plays an important role in mediating the movements of CD-loop.The average MD structure of the actinonin-bound LiPDF complex approaches to the crystal structure.These are consistent with experiment very well. 展开更多
关键词 Peptide deformylase leptospira interrogan Molecular dynamics simulation
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