Prohibitins（PHBs）是高度保守的全能蛋白，因为它们已被证明能介导关键的细胞功能。在这里，我们利用不同的同源模型，表征了编码恶性疟原虫潜在基因的PHBs。我们证明了PfPHB1（PF3D7_0829200）和PfPHB2（PF3D7_1014700）在寄生虫的有性和无性血液阶段都有表达。免疫染色显示这些蛋白质是线粒体驻留蛋白，因为它们被定位为支链结构。我们进一步验证了PfPHB作为器官蛋白质驻留在恶性疟原虫的线粒体中，它们在这里相互作用。在酿酒酵母同源模型中进行了功能特性化，通过在含有相应突变体的细胞中表达PfPHB1和PfPHB2来进行。PfPHBs功能上补充了酵母PHB1和PHB2突变株，这些蛋白质被发现参与稳定线粒体DNA，保持线粒体完整性并拯救酵母细胞生长。此外，已知PHBs和抗癌药物的抑制剂Rocaglamide（Roc-A）被用于检测PfPHBs和作为抗疟药物。 Roc-A处理延缓了PHB1，PHB2和乙啶溴化物小型酵母突变体的生长。此外，Roc-A抑制了用PfPHBs功能性补充的酵母PHB突变体的生长，验证了P. falciparum PHBs作为Roc-A的分子靶点之一。 Roc-A处理以纳摩尔级别抑制青蒿素敏感（3D7），青蒿素耐药（R539T）和氯喹耐药（RKL-9）寄生虫的生长。该化合物能够延缓配子和卵囊的生长，并造成明显的形态异常。基于我们的发现，我们提出了功能性线粒体PfPHB1和PfPHB2在P. falciparum中的存在及其被药物化用于阻止寄生虫生长。版权所有©2023，由Elsevier Inc.出版。

Prohibitins (PHBs) are highly conserved pleiotropic proteins as they have been shown to mediate key cellular functions. Here, we characterize PHBs encoding putative genes of Plasmodium falciparum by exploiting different orthologous models. We demonstrated that PfPHB1 (PF3D7_0829200) and PfPHB2 (PF3D7_1014700) are expressed in asexual and sexual blood stages of the parasite. Immunostaining indicated these proteins as mitochondrial residents as they were found to be localized as branched structures. We further validated PfPHBs as organellar proteins residing in Plasmodium mitochondria, where they interact with each other. Functional characterization was done in Saccharomyces cerevisiae orthologous model by expressing PfPHB1 and PfPHB2 in cells harboring respective mutants. The PfPHBs functionally complemented the yeast PHB1 and PHB2 mutants, where the proteins were found to be involved in stabilizing the mitochondrial DNA, retaining mitochondrial integrity and rescuing yeast cell growth. Further, Rocaglamide (Roc-A), a known inhibitor of PHBs and anti-cancerous agent, was tested against PfPHBs and as an antimalarial. Roc-A treatment retarded the growth of PHB1, PHB2, and ethidium bromide petite yeast mutants. Moreover, Roc-A inhibited growth of yeast PHBs mutants that were functionally complemented with PfPHBs, validating P. falciparum PHBs as one of the molecular targets for Roc-A. Roc-A treatment led to growth inhibition of artemisinin-sensitive (3D7), artemisinin-resistant (R539T) and chloroquine-resistant (RKL-9) parasites in nanomolar ranges. The compound was able to retard gametocyte and oocyst growth with significant morphological aberrations. Based on our findings, we propose the presence of functional mitochondrial PfPHB1 and PfPHB2 in P. falciparum and their druggability to block parasite growth.Copyright © 2023. Published by Elsevier Inc.