Phenoloxidases(POs)are required for the pea aphid s defense against bacterial and fungal infection.Prophenoloxidases(PPOs)are protcolytically converted to its active form PO through a clip domain serine protease casca...Phenoloxidases(POs)are required for the pea aphid s defense against bacterial and fungal infection.Prophenoloxidases(PPOs)are protcolytically converted to its active form PO through a clip domain serine protease cascade.In this study,we identified five clip domain serine proteases in the pea aphids.The messenger RNA levels of two of them,Ap.SPLP and Ap_VP,were upregulated by Gram-positive bacterium Staphylococcus aureus and fungus Beauveha bassiana infections.Double-stranded RNA-based expression knockdown of these two genes resulted in reduced PO activity of the aphid hemolymph,higher loads of S.aureus and B.bassiana in the aphids,and lower survival rates of the aphids after infections.Our data suggest that Ap.SPLP and Ap_VP are involved in PPO activation pathway in the pea aphid.展开更多
Human blood coagulation factor XI （FXI） is a key enzyme in the amplification phase of blood coagulation cascade, and is recognized as an important target for anti-coagulant development in recent years. We designed a...Human blood coagulation factor XI （FXI） is a key enzyme in the amplification phase of blood coagulation cascade, and is recognized as an important target for anti-coagulant development in recent years. We designed a new mutant form of FXIa catalytic domain rhFXI370-607 （N73Q-N113Q-C123S）, and report here the facile preparation, protein crystallization, and crystal structure of this protein. We highlight a few unique structural features of FXIa after comparison with the trypsin family serine proteases at sequence and structural levels. This work provides a foundation to develop new small molecular FXIa inhibitors with increased potency and specificity.展开更多
目的:探讨HtrA1(high temperature requirement A1)在子痫前期孕妇血清、胎盘组织中的表达及意义。方法:HtrA1属分泌型蛋白,可采用酶联免疫吸附法(ELISA)测定同期住院孕妇的空腹血清HtrA1浓度,其中正常晚期妊娠30例、轻度子痫前期25例...目的:探讨HtrA1(high temperature requirement A1)在子痫前期孕妇血清、胎盘组织中的表达及意义。方法:HtrA1属分泌型蛋白,可采用酶联免疫吸附法(ELISA)测定同期住院孕妇的空腹血清HtrA1浓度,其中正常晚期妊娠30例、轻度子痫前期25例、重度子痫前期20例;同时应用免疫组化SABC法检测胎盘组织中HtrA1的表达。结果:正常晚期妊娠、轻度子痫前期及重度子痫前期孕妇血清HtrA1浓度分别为(160.63±32.52)pg/ml、(210.82±32.52)pg/ml及(233.68±38.61)pg/ml,组间差异显著(P<0.05);正常晚期妊娠、轻度子痫前期及重度子痫前期孕妇胎盘组织中HtrA的平均灰度值分别为156.03±5.07、149.89±2.69及140.05±4.96,组间差异显著(P<0.05)。结论:随着病情加重,孕妇血清及胎盘组织中HtrA1的表达显著增高。HtrA1与子痫前期的发生、发展有关。展开更多
Aberrant functioning of serine proteases in inflammatory and carcinogenic processes within the gastrointestinal tract(GIT)has prompted scientists to investigate the potential of serine protease inhibitors,both natural...Aberrant functioning of serine proteases in inflammatory and carcinogenic processes within the gastrointestinal tract(GIT)has prompted scientists to investigate the potential of serine protease inhibitors,both natural and synthetic,as modulators of their proteolytic activities.Protease inhibitors of the Bowman-Birk type,a major protease inhibitor family in legume seeds,which inhibit potently and specifically trypsin-and chymotrypsin-like proteases,are currently being investigated as colorectal chemopreventive agents.Physiologically relevant amounts of Bowman-Birk inhibitors(BBI)can reach the large intestine in active form due to their extraordinary resistance to extreme conditions within the GIT.Studies in animal models have proven that dietary BBI from several legume sources,including soybean,pea,lentil and chickpea,can prevent or suppress carcinogenic and inflammatory processes within the GIT.Although the therapeutic targets and the action mechanism of BBI have not yet been elucidated,the emerging evidence suggests that BBI exert their preventive properties via protease inhibition;in this sense,serine proteases should be considered as primary targets in early stages of carcinogenesis.The validation of candidate serine proteases as therapeutic targets together with the identification,within the wide array of natural BBI variants,of the most potent and specific protease inhibitors,are necessary to better understand the potential of this protein family as colorectal chemopreventive agents.展开更多
基金This work was financially supported by the National Natural Science Foundation of China grant 31772530Natural Science Foundation of Shaanxi Province(2017JM3003)to ZL.
文摘Phenoloxidases(POs)are required for the pea aphid s defense against bacterial and fungal infection.Prophenoloxidases(PPOs)are protcolytically converted to its active form PO through a clip domain serine protease cascade.In this study,we identified five clip domain serine proteases in the pea aphids.The messenger RNA levels of two of them,Ap.SPLP and Ap_VP,were upregulated by Gram-positive bacterium Staphylococcus aureus and fungus Beauveha bassiana infections.Double-stranded RNA-based expression knockdown of these two genes resulted in reduced PO activity of the aphid hemolymph,higher loads of S.aureus and B.bassiana in the aphids,and lower survival rates of the aphids after infections.Our data suggest that Ap.SPLP and Ap_VP are involved in PPO activation pathway in the pea aphid.
基金Supported by FJIRSM (SZD08003)the National Natural Science Foundation of China(30811130467,30625011,30770429)Chinese Academy of Sciences (KSCX2-YW-R-082)
文摘Human blood coagulation factor XI （FXI） is a key enzyme in the amplification phase of blood coagulation cascade, and is recognized as an important target for anti-coagulant development in recent years. We designed a new mutant form of FXIa catalytic domain rhFXI370-607 （N73Q-N113Q-C123S）, and report here the facile preparation, protein crystallization, and crystal structure of this protein. We highlight a few unique structural features of FXIa after comparison with the trypsin family serine proteases at sequence and structural levels. This work provides a foundation to develop new small molecular FXIa inhibitors with increased potency and specificity.
文摘目的:探讨HtrA1(high temperature requirement A1)在子痫前期孕妇血清、胎盘组织中的表达及意义。方法:HtrA1属分泌型蛋白,可采用酶联免疫吸附法(ELISA)测定同期住院孕妇的空腹血清HtrA1浓度,其中正常晚期妊娠30例、轻度子痫前期25例、重度子痫前期20例;同时应用免疫组化SABC法检测胎盘组织中HtrA1的表达。结果:正常晚期妊娠、轻度子痫前期及重度子痫前期孕妇血清HtrA1浓度分别为(160.63±32.52)pg/ml、(210.82±32.52)pg/ml及(233.68±38.61)pg/ml,组间差异显著(P<0.05);正常晚期妊娠、轻度子痫前期及重度子痫前期孕妇胎盘组织中HtrA的平均灰度值分别为156.03±5.07、149.89±2.69及140.05±4.96,组间差异显著(P<0.05)。结论:随着病情加重,孕妇血清及胎盘组织中HtrA1的表达显著增高。HtrA1与子痫前期的发生、发展有关。
基金Supported by ERDF-co-financed grant from the Spanish CICYT,No.AGL2011-26353 to Clemente AClemente A involved in COST Action FA1005 INFOGEST on Food Digestion
文摘Aberrant functioning of serine proteases in inflammatory and carcinogenic processes within the gastrointestinal tract(GIT)has prompted scientists to investigate the potential of serine protease inhibitors,both natural and synthetic,as modulators of their proteolytic activities.Protease inhibitors of the Bowman-Birk type,a major protease inhibitor family in legume seeds,which inhibit potently and specifically trypsin-and chymotrypsin-like proteases,are currently being investigated as colorectal chemopreventive agents.Physiologically relevant amounts of Bowman-Birk inhibitors(BBI)can reach the large intestine in active form due to their extraordinary resistance to extreme conditions within the GIT.Studies in animal models have proven that dietary BBI from several legume sources,including soybean,pea,lentil and chickpea,can prevent or suppress carcinogenic and inflammatory processes within the GIT.Although the therapeutic targets and the action mechanism of BBI have not yet been elucidated,the emerging evidence suggests that BBI exert their preventive properties via protease inhibition;in this sense,serine proteases should be considered as primary targets in early stages of carcinogenesis.The validation of candidate serine proteases as therapeutic targets together with the identification,within the wide array of natural BBI variants,of the most potent and specific protease inhibitors,are necessary to better understand the potential of this protein family as colorectal chemopreventive agents.
