Expression and biochemical characterization of the putative insulinase enzyme PF11_0189 found in the Plasmodium falciparum genome

IF 1.4 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS Protein expression and purification Pub Date : 2024-07-01 DOI:10.1016/j.pep.2024.106539

Prabhash Jyoti Mahanta, Kimjolly Lhouvum

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Abstract

PF11_0189 is a putative insulin degrading enzyme present in Plasmodium falciparum genome. The catalytic domain of PF11_0189 is about 27 kDa. Substrate specificity study shows PF11_0189 acts upon different types of proteins. The substrate specificity is found to be highest when insulin is used as a substrate. Metal dependency study shows highest dependency of PF11_0189 towards zinc metal for its proteolytic activity. Chelation of zinc metal with EDTA shows complete absence of PF11_0189 activity. Peptide inhibitors, P-70 and P-121 from combinatorial peptide library prepared against PF11_0189 show inhibition with an IC50 value of 4.8 μM and 7.5 μM respectively. A proven natural anti-malarial peptide cyclosporin A shows complete inhibition against PF11_0189 with an IC50 value of 0.75 μM suggesting PF11_0189 as a potential target for peptide inhibitors. The study implicates that PF11_0189 is a zinc metalloprotease involved in catalysis of insulin. The study gives a preliminary insight into the mechanism of complications arising from glucose abnormalities during severe malaria.

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恶性疟原虫基因组中发现的假定胰岛素酶 PF11_0189 的表达和生化特征。

PF11_0189 是恶性疟原虫基因组中的一种假定的胰岛素降解酶。PF11_0189 的催化结构域约为 27kDa。底物特异性研究表明，PF11_0189 可作用于不同类型的蛋白质。以胰岛素为底物时，底物特异性最高。金属依赖性研究表明，PF11_0189 的蛋白水解活性对金属锌的依赖性最高。用乙二胺四乙酸螯合锌金属后，PF11_0189 的活性完全消失。从组合肽库中制备的肽抑制剂 P-70 和 P-121 对 PF11_0189 有抑制作用，其 IC50 值分别为 4.8 μM 和 7.5 μM。一种已被证实的天然抗疟疾肽环孢素 A 对 PF11_0189 显示出完全的抑制作用，IC50 值为 0.75 μM，这表明 PF11_0189 是肽抑制剂的一个潜在靶点。该研究表明，PF11_0189 是一种参与催化胰岛素的锌金属蛋白酶。该研究初步揭示了严重疟疾期间葡萄糖异常引起并发症的机制。

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来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.