Objective:To identify a full length cDNA sequence of a novel tetraspanin(TSP) homologue from Spirometra erinaceieuropaei and to predict the structure and function of its encoding protein using bioinformatics methods.M...Objective:To identify a full length cDNA sequence of a novel tetraspanin(TSP) homologue from Spirometra erinaceieuropaei and to predict the structure and function of its encoding protein using bioinformatics methods.Methods:Using the NCBI,EMBI,Expasy and other online sites, the open reading frame(ORF),conserved domain,physical and chemical parameters,signal peptide,transmembrane domain,epitope,topological structures of the protein sequences were predicted.And Vector NTI software was used for multiple sequence alignment and phylogenetic tree construction.Results:’Hie target sequence was 1 132 hp length with a 681 hp biggest ORF encoding 226 amino acids protein with typical TSP conserved domain.It was confirmed as full length cDNA of TSP16 from Spirometra erinaceieuropaei and named as SeTSP16 (GenBank accession number:JF728872).The predicted molecular weight and isoelectric point of the deduced protein were 24 750.5 Da and 7.88 Da,respectively.Compared with TSP16s from Schistosoma japonicum and Schistosoma mansoni.it showed similarity of 59%and 59%, respectively.SeTSP16 contained four transmembrane domains(TM 1-4),intracellular N and C-termini,one short small extracellular loop and one large extracellular loop.Four major epitopes that were significant different from the corresponding epitope regions of TSP16 from Schistosoma mansoni and Schistosoma japonicum were predicted.Conclusions:The full length cDNA sequences of SeTSP16 arc identified.It encodes a transmembrane protein which might be an ideal diagnosis antigen and target molecule for antiparasitic drugs.展开更多
This study characterized the activation of platelet integrin α bβ3 induced by two anti human platelet tetraspanin monoclonal antibodies(mAbs),HI117 and SJ9A4. Methods.Using 125 I labeled human fibrinogen(Fg),specifi...This study characterized the activation of platelet integrin α bβ3 induced by two anti human platelet tetraspanin monoclonal antibodies(mAbs),HI117 and SJ9A4. Methods.Using 125 I labeled human fibrinogen(Fg),specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured as indication of activation of platelet integrin αbβ3 by the two mAbs. Results.HI117 and SJ9A4(10μg/ml and 20μg/ml) induced evident specific Fg binding to human platelets,suggesting that the two mAbs evoked activation of platelet integrin αbβ3.Further study indicated that HI117 and SJ9A4 induced integrin αⅡbβ3 activation independent of platelet Fc receptors, and that HI117 and SJ9A4 induced integrin αbβ3 activation was inhibited by sphingosing, aspirin, apyrase, and/or PGI2. Conclusion.The anti platelet tetraspanin(CD9)mAbs,HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 activation independent of Fc receptors.Three signaling pathways,i.e.thromboxane,secreted ADP, and cAMP pathways may be involved in the process,with protein kinase C activation presumably being the common step of the three pathways.展开更多
characterize the activation of platelet integrin αⅡbβ3 induced by two anti human platelet tetraspanin monoclonal antibodies (mAbs), HI117 and SJ9A4, and investigate their potential mechanism of action Method...characterize the activation of platelet integrin αⅡbβ3 induced by two anti human platelet tetraspanin monoclonal antibodies (mAbs), HI117 and SJ9A4, and investigate their potential mechanism of action Methods Using 125 I labeled human fibrinogen (Fg), specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured Results HI117 and SJ9A4 (10?μg/ml and 20?μg/ml) induced specific Fg binding to human platelets, suggesting that the two mAbs evoked activation of platelet integrin αⅡbβ3 Further study indicated that HI117 and SJ9A4 induced integrin αⅡbβ3 activation independent of platelet Fc receptors, and that HI117 and SJ9A4 induced integrin αⅡbβ3 activation was inhibited by pretreatment of platelets with sphingosine, aspirin, apyrase, and/or PGI 2 Conclusions Anti platelet tetraspanin (CD9) mAbs, HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 activation independent of Fc receptors Three signaling pathways, namely thromboxane, secreted ADP, and cAMP pathways, may be involved in the process, with protein kinase C activation presumably being the common step of the three pathways展开更多
文摘Objective:To identify a full length cDNA sequence of a novel tetraspanin(TSP) homologue from Spirometra erinaceieuropaei and to predict the structure and function of its encoding protein using bioinformatics methods.Methods:Using the NCBI,EMBI,Expasy and other online sites, the open reading frame(ORF),conserved domain,physical and chemical parameters,signal peptide,transmembrane domain,epitope,topological structures of the protein sequences were predicted.And Vector NTI software was used for multiple sequence alignment and phylogenetic tree construction.Results:’Hie target sequence was 1 132 hp length with a 681 hp biggest ORF encoding 226 amino acids protein with typical TSP conserved domain.It was confirmed as full length cDNA of TSP16 from Spirometra erinaceieuropaei and named as SeTSP16 (GenBank accession number:JF728872).The predicted molecular weight and isoelectric point of the deduced protein were 24 750.5 Da and 7.88 Da,respectively.Compared with TSP16s from Schistosoma japonicum and Schistosoma mansoni.it showed similarity of 59%and 59%, respectively.SeTSP16 contained four transmembrane domains(TM 1-4),intracellular N and C-termini,one short small extracellular loop and one large extracellular loop.Four major epitopes that were significant different from the corresponding epitope regions of TSP16 from Schistosoma mansoni and Schistosoma japonicum were predicted.Conclusions:The full length cDNA sequences of SeTSP16 arc identified.It encodes a transmembrane protein which might be an ideal diagnosis antigen and target molecule for antiparasitic drugs.
文摘This study characterized the activation of platelet integrin α bβ3 induced by two anti human platelet tetraspanin monoclonal antibodies(mAbs),HI117 and SJ9A4. Methods.Using 125 I labeled human fibrinogen(Fg),specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured as indication of activation of platelet integrin αbβ3 by the two mAbs. Results.HI117 and SJ9A4(10μg/ml and 20μg/ml) induced evident specific Fg binding to human platelets,suggesting that the two mAbs evoked activation of platelet integrin αbβ3.Further study indicated that HI117 and SJ9A4 induced integrin αⅡbβ3 activation independent of platelet Fc receptors, and that HI117 and SJ9A4 induced integrin αbβ3 activation was inhibited by sphingosing, aspirin, apyrase, and/or PGI2. Conclusion.The anti platelet tetraspanin(CD9)mAbs,HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 activation independent of Fc receptors.Three signaling pathways,i.e.thromboxane,secreted ADP, and cAMP pathways may be involved in the process,with protein kinase C activation presumably being the common step of the three pathways.
基金grantfromtheNationalNaturalScienceFoundationofChina (No 39470 172 )
文摘characterize the activation of platelet integrin αⅡbβ3 induced by two anti human platelet tetraspanin monoclonal antibodies (mAbs), HI117 and SJ9A4, and investigate their potential mechanism of action Methods Using 125 I labeled human fibrinogen (Fg), specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured Results HI117 and SJ9A4 (10?μg/ml and 20?μg/ml) induced specific Fg binding to human platelets, suggesting that the two mAbs evoked activation of platelet integrin αⅡbβ3 Further study indicated that HI117 and SJ9A4 induced integrin αⅡbβ3 activation independent of platelet Fc receptors, and that HI117 and SJ9A4 induced integrin αⅡbβ3 activation was inhibited by pretreatment of platelets with sphingosine, aspirin, apyrase, and/or PGI 2 Conclusions Anti platelet tetraspanin (CD9) mAbs, HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 activation independent of Fc receptors Three signaling pathways, namely thromboxane, secreted ADP, and cAMP pathways, may be involved in the process, with protein kinase C activation presumably being the common step of the three pathways