【摘 要】
：
Avian coccidiosis, caused by Eimeria spp., is one of the major parasitic diseases in birds.Cysteine protease is a major virulence factor in parasitic protozoa, and it may be a suitable chemotherapeuti
【机 构】
：
National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University,
【出 处】
：
第七届国际分子模拟与信息技术应用学术会议
论文部分内容阅读
Avian coccidiosis, caused by Eimeria spp., is one of the major parasitic diseases in birds.Cysteine protease is a major virulence factor in parasitic protozoa, and it may be a suitable chemotherapeutic target and vaccine candidate molecule.A 100 amino acid (an.) partial sequence of cathepsin L, which is a cysteine protease, was reported by Katrib et al.(Ac.No.CDJ41293) (2012).A 219 an.sequence was reported by Reid et al.(Ac.No.AFV92863) (2013).However, the open reading frame (ORF) was not reported In this study, a full sequence of a cathepsin-L-like peptidase in Eimeria tenella (EtcatL) was obtained and its biochemical characterizations and expression profiles were analyzed across different stages of the parasites life cycle.Results showed that the EtcatL gene encodes a protein 470 aa.in length, with 47 and 49 % identity to Toxoplasma gondii and Eimeria acervulina.Considering the close phylogenetic relationship, TgcatL (PDB.ID 3F75) was selected for use as a template for homology modeling with quality factors of 90.9.Gelatin SDS-PAGE showed it to exert protease activity at ≈38 and ≈26 kDa.Further analysis showed the kinetic parameters of the recombinant peptidase to be Km-8.9 μM and Vmax--5.7 RFU/s μM at pH 5.5 containing 10 mM dithiothreitol (DTT) in the reaction matrix, and the IC50 value of E64 was 65.32±3.02 nM.The recombinant protein was active from 25 to 50 ℃, with optimal activity at 42 ℃.The RT-PCR and Western blot results showed it to be expressed mainly at the endogenous stages and the initial phase of the sporulation.The protective experiment showed that chickens immunized with 100 and 200 μg rEtcatL had reduction of weight loss values 48.7 and 57.9 % those of infected controls, respectively.Their reduction of lesion scores (RLS) were 25.0 and 47.2 % that of control chickens, and relative oocyst production (ROP) was 39.6 and 15.5 % that of control chickens.These results indicate that the EtcatL can be used as an effective immunogen,and further studies are needed to enhance its potential as a vaccine candidate molecule.
其他文献
Human glutathione transferase zeta 1c-1c (hGSTZ1c-1c) is one of the glutathione transferase isoenzymes and considered to be a protein scaffold to imitate glutathione peroxidase (GPX) owing to the natu
Most lipases contain a lid domain to shield the hydrophobic binding site from the water environment.The lid, mostly in helical form, can undergo a conformational change to expose the active cleft duri
A series of novel indole derivatives was synthesized as inhibitors of fructose-1,6-bisphosphatase (FBPase).Extensive structure-activity relationships were conducted and led to a potent FBPase inhibito
In order to enlarge the substrate binding pocket of the meso-diaminopimelate dehydrogenase from Symbiobacterium thermophilum to accommodate larger 2-keto acids, four amino acid residues (Phe146, Thr17
Targeting the adaptor protein (transforming growth factor-β (TGF-β)-activated protein kinase 1 (TAK1)-binding protein 1) (TAB1)-mediated non-canonical activation of p38α to limit ischemia/reperfusion
Background: Immunotherapy has been used to improve patient immune function, inhibit tumor growth and has become a highly promising method of cancer treatment.Highly agglutinative staphylococcin (HAS),
Both recent studies and our calculation suggest that the physicochemical properties of launched drugs changed continuously over the past decades.Besides shifting of commonly used properties, the avera
Given the high risk and lengthy procedure of traditional drug development, drug repurposing is gaining more and more attention.Although many types of drug information have been used to repurpose drugs
The SNP-158G＞A of KLK3 has been validated as a regulatory SNP (rSNP) by molecular biology assays, but the mechanism of how it affects the binding of an androgen receptor (AR) homodimer with DNA is unc
Motivation: One of the major findings in systems biomedicine is that both pathogenesis of diseases and drug mode of action have a module basis.However, the transcription factors (TFs) regulating the m