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2011年中国水产学会学术年会

作者： Shu Wu1 Shuhong Wang1 Guodong Wang1 Zhihua Zou1 Yilei Wang1 Ziping Zhang2 (1 The Key Laboratory of Science and Technology for Aquaculture and Food Safety,Fisheries College,Jimei University,Xiamen,Fujian 361021,China 2 Department of Biological Science,Seton Hall University,New Jersey,USA,07079) The Key Laboratory of Science and Technology for Aquaculture and Food Safety Fisheries College Jim Department of Biological Science Seton Hall University New Jersey USA 07079

＜正＞Two immune related genes,cysteine protease（sacp） and cysteine proteases inhibitor were cloned by EST analysis、SMART RACE and Genome Walking techniques.Their complete sequences were confirmed by head to toe PCR.Real time PCR was used to analyze their expression patterns in different tissues and the expression time course in hepatopancreas and haemocytes after Vibrio Parahaemolyticus infection.Results are indicated that:The full length cDNA of sacp was 1 558 bp and the deduced protein was composed of 303 amino acids, containing two cysteine protease active sites.BLAST analysis indicated sacp was belong to the type of cathepsin L.The full length cDNA of sacpi was 1 481 bp,the deduced protein was composed of 253 amino acids,containing a characteristic motif of CPI and the structural features of the familyⅡcystatins,consensus sequences of reactive sites:Gly63,Q84VVSG88 and P114W115 pair.Meanwhile,the 5’ flanking sequence of sacpi existed some transcription factor binding sites predicted by software.Tissue distribution and its responses to Vibrio parahaemolyticus challenge also were investigated.

关键词： cysteine protease cysteine proteases inhibitor Haliotis diverscolor

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Ultrastructural and enzymatic alterations in the ovary of Rhodnius prolixus infected with Trypanosoma rangeli

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Asian Pacific Journal of Tropical Medicine 2022年第2期15卷 71-78页

作者： Guilherme S.Machado Rosane L.Lopes Simone S.C.Oliveira Simone P.C.Freitas Jacenir R.Santos-Mallet AndréL.S.Santos Denise Feder Suzete A.O.Gomes Programa de Pós-Graduação em Ciências e Biotecnologia Instituto de BiologiaUniversidade Federal FluminenseNiteróiRJBrazil Laboratório de Biodiversidade de Insetos e Patógenos Departamento de Biologia GeralInstituto de BiologiaUniversidade Federal FluminenseNiteróiRJBrazil Laboratório de Estudos Avançados de Microrganismos Emergentes e Resistentes Departamento de Microbiologia GeralInstituto de Microbiologia Paulo de GóesUniversidade Federal do Rio de JaneiroRJBrazil Fundação Oswaldo Cruz Piauí TeresinaPIBrazil Laboratório Interdisciplinar de Vigilância Entomológica em Díptera e Hemíptera Instituto Oswaldo CruzFundação Oswaldo CruzRio de JaneiroRJBrazil Universidade Iguaçu Nova IguaçuRJBrazil Laboratório de Biologia de Insetos Departamento de Biologia GeralInstituto de BiologiaUniversidade Federal FluminenseNiteróiRJBrazil

Objective:To investigate the morphological structure of ovarian follicular cells and biochemical parameters of both ovaries and fat bodies(sites of vitellogenesis)from Rhodnius(R.)prolixus infected with Trypanosoma(T.)rangeli.Methods:Adult virgin females of R.prolixus were fed upon a membrane apparatus containing heat-inactivated citrated rabbit blood and a suspension of T.rangeli epimastigotes(Macias strain).Females from the control group and all the males received parasite-free blood.Transmission electron microscopy was used to reveal the morphological aspects of ovarian follicle cells in both control and parasite-infected groups.Protein profile,proteolytic activities and Western blotting analyses were performed in either ovary or fat body samples of control and parasite-infected groups.Results:According to the ultrastructural data,T.rangeli infection elicited a degeneration process in the ovarian follicular cells of R.prolixus.Proteolytic assays indicated a reduction in the activity of aspartic peptidases in the ovary and fat body from parasite-infected group,while a significant increase in the cysteine peptidase activity was measured in both insect organs.Additionally,immunoblotting revealed that vitellogenin was overexpressed in the ovary of parasite-infected insects.Conclusions:T.rangeli infection seems to elicit an early programmed cell death in the ovarian follicle cells as well as induces the modulation on the activities of different peptidase classes in either ovaries or fat bodies and the overexpression of the vitellogenin in the ovary of R.prolixus.

关键词： Trypanosoma rangeli Rhodnius prolixus Ovarian follicle Vitellogenin Aspartic protease cysteine protease

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cysteine proteases as therapeutic targets:does selectivity matter? A systematic review of calpain and cathepsin inhibitors

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Acta Pharmaceutica Sinica B 2015年第6期5卷 506-519页

作者： Marton Siklos Manel Ben Aissa Gregory R.J.Thatcher Department of Medicinal Chemistry and Pharmacognosy University of Illinois College of Pharmacy

cysteine proteases continue to provide validated targets for treatment of human diseases.In neurodegenerative disorders,multiple cysteine proteases provide targets for enzyme inhibitors,notably caspases,calpains,and cathepsins.The reactive,active-site cysteine provides specificity for many inhibitor designs over other families of proteases,such as aspartate and serine;however,a)inhibitor strategies often use covalent enzyme modification,and b)obtaining selectivity within families of cysteine proteases and their isozymes is problematic.This review provides a general update on strategies for cysteine protease inhibitor design and a focus on cathepsin B and calpain 1 as drug targets for neurodegenerative disorders;the latter focus providing an interesting query for the contemporary assumptions that irreversible,covalent protein modification and low selectivity are anathema to therapeutic safety and efficacy.

关键词： cysteine protease Calpain Cathepsin Enzyme inhibitors Neurodegeneration Alzheimer’s disease

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Molecular mechanisms of insect adaptation to plant defense： Lessons learned from a Bruchid beetle

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Insect Science 2008年第6期15卷 477-481页

作者： Keyan Zhu-Salzman Ren Sen Zeng Department of Entomology Texas A&M University College Station Texas USA Vegetable & Fruit Improvement Center Texas A&M University College Station Texas USA Institute of Tropical and Subtropical Ecology South China Agricultural University Guangzhou China

Plants can accumulate, constitutively and/or after induction, a wide variety of defense compounds in their tissues that confer resistance to herbivorous insects. The naturally occurring plant resistance gene pool can serve as an arsenal in pest management via transgenic approaches. As insect-plant interaction research rapidly advances, it has gradually become clear that the effects of plant defense compounds are determined not only by their toxicity toward target sites, but also by how insects respond to the challenge. Insect digestive tracts are not passive targets of plant defense, but often can adapt to dietary challenge and successfully deal with various plant toxins and anti-metabolites. This adaptive response has posed an obstacle to biotechnology-based pest control approaches, which underscores the importance of understanding insect adaptive mechanisms. Molecular studies on the impact of protease inhibitors on insect digestion have contributed significantly to our understanding of insect adaptation to plant defense. This review will focus on exposing how the insect responds to protease inhibitors by both qualitative and quantitative remodeling of their digestive proteases using the cowpea bruchid-soybean cysteine protease inhibitor N system.

关键词： adaptation cowpea bruchid cysteine protease gene regulation genomics protease inhibitor scN

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Effects of nerve growth factor on the expression of caspase-12 of nerve cells in cerebral ischemia/reperfusion area

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Neural Regeneration Research 2006年第4期1卷 293-296页

作者： Jiping Yang Huaijun Liu Ying Li Yan Liu Haiqing Yang Department of Medical Imaging Second Hospital Hebei Medical University Shijiazhuang 050000 Hebei Province China Department of Internal Medicine Second Hospital Hebei Medical University Shijiazhuang 050000 Hebei Province China

BACKGROUND： Researches suggest that cascade reaction of cysteine protease mediated by caspase-12 can cause apoptosis after cerebral ischemia/reperfusion injury; however, nerve growth factor （NGF） can reduce apoptosis through inhibiting activation of that reaction. OBJECTIVE： To observe the effect of NGF on the expression of caspase-12 in brain tissue of rabbits with cerebral ischemia/reperfusion injury, and elucidate the protective mechanism of NGF on neural apoptosis induced by cerebral ischemia/reperfusion injury. DESIGN： Randomized controlled animal study SETTING： Department of Image, Second Hospital, Hebei Medical University .MATERIALS ： A total of 26 healthy New Zealand rabbits, of clean grade, aged 4.5-5 months, weighing （2.6±0.2） kg, were selected in this study. Reagents： NGF （Xiamen Beida Zhilu Biotechnology Co., Ltd.）; caspase-12 （Santa Cruz Biotechnology Company, USA, clone number： SC-12395）; caspase-3 （Santa Cruz Biotechnology Company, USA, clone number： SC-7272）; biotin-antibody Ⅱ and ABC compound （Wuhan Boster Company）; in situ end-labeling （ISEL, Beijing Zhongshan Company）. METHODS： The experiment was carried out in the Laboratories of Nerve Molecule Image Science and Neurology of the Second Hospital of Hebei Medical University from May to August 2005. （1） All animals were randomly divided into three groups. Ischemia/reperfusion （I/R） group （n=10）： Left middle cerebral artery （MCA） was blocked for 2 hours and then blooded for 2 hours in order to establish focal cerebral ischemi- a/reperfusion models. Sham operation group （n=6）： Cork was inserted with 3 cm in depth, and then pulled to common carotid artery. Other procedures were as the same as those in ischemia/reperfusion group. Treatment group （n=-10）： After modeling, 400 AU （16 μg/L） NGF was inserted into cerebral infarction focus immediately. （2） Brain tissue was taken out from left ischemia/reperfusion area at 3 days after operation in sham operation group and at 3 days after reperfusion in other two groups, In addition, contents of ca

关键词： focal cerebral ischemia nerve cells cascade reaction middle cerebral artery common carotid artery cerebral infarction cysteine protease flow cytometry growth factor animal study New Zealand in order

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Identification of Molecular Marker Linked to Salt Tolerance Gene in Alfalfa

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Agricultural Sciences in China 2005年第10期4卷 781-787页

作者： YANG Qing-chuan HAN Jian-guo SUN Yan WU Ming-sheng Institute of Animal Science Chinese Academy of Agricultural Sciences Beijing 100094 R.P.China Institute of Grassland Science China Agricultural University Beijing 100094 R.P.China

The study has established the F2 offspring obtained by crossing salt-tolerant with salt-sensitive alfalfa, and appraised the salt-tolerant F2 offspring seedling was evaluated in pot culture. With the F2 segregated population, the research has obtained a molecular marker linked with salt-tolerant genes of alfalfa using the improved BSA combined with RAPD. The RAPD PCR products were excised from the agarose gel and purified using a kit, then were mixed with pMD-18T vector and sequenced. Sequencing result indicated the RAPD marker was 1 438 bp in length. Similarity researches using blast in Genbank indicated that the nucleotide sequence of the RAPD marker showed 93% and 91% similarity with mth2-6el8 gene fragment （347 bp） and ruth2-33122 gene fragment （334 bp） of Medicago truncatula respectively. Medicago truncatula is a close relative of alfalfa and Mth2-6el8 is a molecular marker of the gene coding for a cysteine protease which was saltinducible in some plants. These results indicated the RAPD marker was possibly related to cysteine protease genes in alfalfa.

关键词： Alfalfa Salt tolerance Molecular marker cysteine protease

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Identification and characterization of a cathepsin-F-like protease in Trichinella spiralis

Identification and characterization of a cathepsin-F-like pr...

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中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

作者： Zi-gang Qu Xue-ting Ma Wen-hui Li Nian-zhang Zhang Long Yue Jian-min Cui Jian-ping Cai Wan-zhong Jia Bao-quan Fu State Key Laboratory of Veterinary Etiological Biology Key Laboratory of Veterinary Public Health of the Ministry of AgricultureKey Laboratory of Veterinary Parasitology of Gansu ProvinceLanzhou Veterinary Research InstituteChinese Academy of Agricultural Sciences Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease

Trichinellosis is a an emerging and reemerging infectious disease,caused by Trichinella spp.Cathepsin F belongs to cysteine protease that is a major virulence factor for parasitic helminths,and it may be a potential anti-helminth drug target and vaccine candidate.In this study,the cDNA encoding a putative cathepsin F-like peptidase in Trichinella spiralis,TsCF1,was obtained and its biochemical characterization and expression profile were analyzed.Results demonstrated that the TsCF1 gene encodes a protein of 366 aa with the theoretical molecular weight of 41.9 ku and isoelectric point of 7.46.The signal peptide sequence of TsCF1 located between amino acids 1 and 19 and 3 putative N-glycosylation sites were predicted.The cysteine protease conserved active site of Cys173,His309 and Asn333 were identified and cathepsin F specific motif ERFNAQ like KLFNAQ sequence was revealed in the propeptide of TsCF1.Homology analysis revealed a higher than 40%identity with other cathepsin F from parasitic helminth and phylogenetic analysis indicated TsCF1 located in the different clans with trematode cathepsin Fs.RT-PCR revealed the gene was expressed in muscle larvae,newborn larvae and adult stages.Gelatin SDS-PAGE revealed the recombinant protein was expressed with the molecular weight of 45 ku.Immunolocalization analysis showed that TsCF1 located on the inner layer of the cuticle and stichosome of the parasite,which probably play a vital role in nutrition uptake.The purified recombinant TsCF1 was used to immunize rabbit and the immune serum could recognize a band of soluble protein of muscle larvae at about 46 ku.After renaturation the TsCF1 recombinat protein demonstrated substantial enzyme activity to Z-Phe-Arg-AMC substrate with the optimal pH 5.5 and this activity could be inhibited by cysteine protease inhibitor E-64.Further analysis revealed the kinetic parameters of the recombinant peptidase to be Km=0.5091 μM and Vmax= 6.12 RFU/sμM at pH 5.5,and the IC50 value of E64 was 135.50±16.90 nM.These results indicated that the

关键词： Trichinella spiralis cysteine protease Cathepsin F enzyme activity

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