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首页> 外文期刊>Journal of Agricultural and Food Chemistry >AN IN VITRO ASSAY FOR MONITORING PRENYL TRANSFERASE ACTIVITY IN LEPIDOPTERAN CORPORA ALLATA
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AN IN VITRO ASSAY FOR MONITORING PRENYL TRANSFERASE ACTIVITY IN LEPIDOPTERAN CORPORA ALLATA

机译：监测斜纹夜蛾异戊二烯转移酶活性的体外测定

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摘要

The prenyl transferase, farnesyl pyrophosphate (FPP) synthase, which catalyzes the head-to-tail couplings of dimethylallyl pyrophosphate (DMAPP) and geranyl pyrophosphate (GPP) with isopentenyl pyrophosphate (IPP), is a key enzyme in juvenile hormone (JH) biosynthesis. While moderate FPP synthase activity has been found in whole-body insect preparations, enzymatic activity in the corpus allatum, the gland responsible for JH biosynthesis, has been elusive. A new procedure for examining corpora allata prenyl transferase activity in the lepidopteran Manduca sexta is described. In contrast to most other short-chain prenyl transferases, the larval enzyme requires detergent for activity. Optimum conversion of DMAPP and GPP to FPP is obtained with corpora allata homogenates containing 2% Triton X-100, 0.15% bovine serum albumin, and 25% glycerol. The enzyme is activated in the presence of Mn2+ or Mg2+ and is inactivated by the addition of N-ethylmaleimide.
机译：异戊烯基焦磷酸（FPP）合酶异戊烯基转移酶催化二甲基烯丙基焦磷酸（DMAPP）和香叶基焦磷酸（GPP）与异戊烯基焦磷酸（IPP）的头尾耦合，是少年激素（JH）生物合成中的关键酶。 。尽管在全身昆虫制剂中发现了适度的FPP合酶活性，但难以捉摸的负责JH生物合成的腺体-黄体中的酶促活性。描述了一种新的程序，用于检查鳞翅目满天蛾六倍体中的全集变应原异戊二烯基转移酶活性。与大多数其他短链异戊二烯基转移酶相反，幼虫酶需要去污剂才能发挥活性。使用含2％Triton X-100、0.15％牛血清白蛋白和25％甘油的全集匀浆，可以将DMAPP和GPP最佳地转化为FPP。该酶在Mn2 +或Mg2 +的存在下被激活，并通过添加N-乙基马来酰亚胺使其失活。

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