Cristian Follmer, Rafael Real-Guerra, German E Wasserman, Deiber Olivera-Severo, Célia R Carlini
{"title":"胡豆、大豆和巴氏杀菌杆菌尿素酶:与尿素分解活性无关的生物效应。","authors":"Cristian Follmer, Rafael Real-Guerra, German E Wasserman, Deiber Olivera-Severo, Célia R Carlini","doi":"10.1111/j.1432-1033.2004.04046.x","DOIUrl":null,"url":null,"abstract":"<p><p>In this work we compared two plant ureases, jackbean urease (JBU) and embryo-specific soybean urease (SBU) and a bacterial (Bacillus pasteurii) urease, for kinetic parameters and other biological properties described recently for ureases that are independent of the ureolytic activity. The insecticidal effect of ureases was investigated in feeding trials with the cotton sucker bug, Dysdercus peruvianus (Hemiptera) as an insect model. Contrasting with B. pasteurii urease (PBU), both plant ureases presented potent insecticidal activity, with LD(50) values of 0.017% (w/w) and 0.052% (w/w) for JBU and SBU, respectively. The insecticidal property of JBU or SBU was not affected by treatment with p-hydroxymercuribenzoate, an irreversible inhibitor of ureolytic activity of both proteins. Also, contrasting with canatoxin - a urease isoform from jackbean seeds that displays a toxic effect in mice (LD(50) = 2 mg x kg(-1)) - no lethality was seen in mice injected intraperitoneally with JBU or SBU (20 mg x kg(-1)). Similarly to canatoxin, the three enzymes promoted aggregation of blood platelets (EC(50) = 400.0 micro g x mL(-1), 22.2 micro g x mL(-1), 15.8 micro g x mL(-1) for BPU, SBU and JBU, respectively). This platelet activating property was also independent of urease activity. Comparison of the kinetic properties indicated that SBU is fivefold less susceptible than JBU to inhibition by acetohydroxamic acid, a chelator of Ni(+2) and Zn(+2) ions. The ureases also showed different susceptibility to agents that modify cysteine residues, such as p-hydroxymercuribenzoate and p-benzoquinone. Altogether, these data emphasize that biological properties that are independent of ureolytic activity are not restricted to jackbean ureases and that these proteins may have a role in plant defense against insect predators.</p>","PeriodicalId":11817,"journal":{"name":"European journal of biochemistry","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2004-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/j.1432-1033.2004.04046.x","citationCount":"99","resultStr":"{\"title\":\"Jackbean, soybean and Bacillus pasteurii ureases: biological effects unrelated to ureolytic activity.\",\"authors\":\"Cristian Follmer, Rafael Real-Guerra, German E Wasserman, Deiber Olivera-Severo, Célia R Carlini\",\"doi\":\"10.1111/j.1432-1033.2004.04046.x\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>In this work we compared two plant ureases, jackbean urease (JBU) and embryo-specific soybean urease (SBU) and a bacterial (Bacillus pasteurii) urease, for kinetic parameters and other biological properties described recently for ureases that are independent of the ureolytic activity. The insecticidal effect of ureases was investigated in feeding trials with the cotton sucker bug, Dysdercus peruvianus (Hemiptera) as an insect model. Contrasting with B. pasteurii urease (PBU), both plant ureases presented potent insecticidal activity, with LD(50) values of 0.017% (w/w) and 0.052% (w/w) for JBU and SBU, respectively. The insecticidal property of JBU or SBU was not affected by treatment with p-hydroxymercuribenzoate, an irreversible inhibitor of ureolytic activity of both proteins. Also, contrasting with canatoxin - a urease isoform from jackbean seeds that displays a toxic effect in mice (LD(50) = 2 mg x kg(-1)) - no lethality was seen in mice injected intraperitoneally with JBU or SBU (20 mg x kg(-1)). Similarly to canatoxin, the three enzymes promoted aggregation of blood platelets (EC(50) = 400.0 micro g x mL(-1), 22.2 micro g x mL(-1), 15.8 micro g x mL(-1) for BPU, SBU and JBU, respectively). This platelet activating property was also independent of urease activity. Comparison of the kinetic properties indicated that SBU is fivefold less susceptible than JBU to inhibition by acetohydroxamic acid, a chelator of Ni(+2) and Zn(+2) ions. The ureases also showed different susceptibility to agents that modify cysteine residues, such as p-hydroxymercuribenzoate and p-benzoquinone. Altogether, these data emphasize that biological properties that are independent of ureolytic activity are not restricted to jackbean ureases and that these proteins may have a role in plant defense against insect predators.</p>\",\"PeriodicalId\":11817,\"journal\":{\"name\":\"European journal of biochemistry\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2004-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1111/j.1432-1033.2004.04046.x\",\"citationCount\":\"99\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"European journal of biochemistry\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1111/j.1432-1033.2004.04046.x\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"European journal of biochemistry","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1111/j.1432-1033.2004.04046.x","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 99
摘要
在这项工作中,我们比较了两种植物脲酶(大豆脲酶(JBU)和胚特异性大豆脲酶(SBU))和一种细菌(巴氏芽孢杆菌)脲酶的动力学参数和最近描述的与脲酶分解活性无关的其他生物特性。在以棉花吸盘虫 Dysdercus peruvianus(半翅目)为昆虫模型的饲养试验中,研究了脲酶的杀虫效果。与巴氏杆菌脲酶(PBU)相比,两种植物脲酶都具有很强的杀虫活性,JBU 和 SBU 的 LD(50) 值分别为 0.017%(重量/重量)和 0.052%(重量/重量)。JBU 或 SBU 的杀虫特性不受对羟基脲苯甲酸酯处理的影响,对羟基脲苯甲酸酯是这两种蛋白质尿解活性的不可逆抑制剂。此外,与小鼠腹腔注射 JBU 或 SBU(20 毫克 x 千克(-1))不会导致小鼠死亡的卡纳毒素相反,小鼠腹腔注射 JBU 或 SBU(20 毫克 x 千克(-1))不会导致小鼠死亡。与卡纳毒素相似,这三种酶也能促进血小板聚集(BPU、SBU 和 JBU 的 EC(50) = 400.0 微克 x 毫升(-1)、22.2 微克 x 毫升(-1)、15.8 微克 x 毫升(-1))。这种血小板活化特性也与脲酶活性无关。动力学特性的比较表明,SBU 受乙酰羟肟酸(一种镍(+2)和锌(+2)离子的螯合剂)抑制的敏感性比 JBU 低五倍。脲酶对修饰半胱氨酸残基的制剂(如对羟基巯基苯甲酸酯和对苯醌)的敏感性也有所不同。总之,这些数据强调了独立于尿素分解活性的生物特性并不局限于胡豆尿素酶,而且这些蛋白质可能在植物抵御昆虫天敌的过程中发挥作用。
Jackbean, soybean and Bacillus pasteurii ureases: biological effects unrelated to ureolytic activity.
In this work we compared two plant ureases, jackbean urease (JBU) and embryo-specific soybean urease (SBU) and a bacterial (Bacillus pasteurii) urease, for kinetic parameters and other biological properties described recently for ureases that are independent of the ureolytic activity. The insecticidal effect of ureases was investigated in feeding trials with the cotton sucker bug, Dysdercus peruvianus (Hemiptera) as an insect model. Contrasting with B. pasteurii urease (PBU), both plant ureases presented potent insecticidal activity, with LD(50) values of 0.017% (w/w) and 0.052% (w/w) for JBU and SBU, respectively. The insecticidal property of JBU or SBU was not affected by treatment with p-hydroxymercuribenzoate, an irreversible inhibitor of ureolytic activity of both proteins. Also, contrasting with canatoxin - a urease isoform from jackbean seeds that displays a toxic effect in mice (LD(50) = 2 mg x kg(-1)) - no lethality was seen in mice injected intraperitoneally with JBU or SBU (20 mg x kg(-1)). Similarly to canatoxin, the three enzymes promoted aggregation of blood platelets (EC(50) = 400.0 micro g x mL(-1), 22.2 micro g x mL(-1), 15.8 micro g x mL(-1) for BPU, SBU and JBU, respectively). This platelet activating property was also independent of urease activity. Comparison of the kinetic properties indicated that SBU is fivefold less susceptible than JBU to inhibition by acetohydroxamic acid, a chelator of Ni(+2) and Zn(+2) ions. The ureases also showed different susceptibility to agents that modify cysteine residues, such as p-hydroxymercuribenzoate and p-benzoquinone. Altogether, these data emphasize that biological properties that are independent of ureolytic activity are not restricted to jackbean ureases and that these proteins may have a role in plant defense against insect predators.