The Aga Khan University, Karachi, Pakistan.
Chem Biol Drug Des. 2019 Mar;93(3):351-363. doi: 10.1111/cbdd.13421. Epub 2019 Jan 29.
The evolution of voltage-gated calcium channel (Cav) in eukaryotes is an area of interest for biologists worldwide. The CLAN CL0030 and its family Ion_Trans 2 PF 07885 have been known to be present in prokaryotes, but the origin of these ion channels in Acanthamoeba spp. is yet to be determined. We inferred the origin of primitive forms of two-pore channels like proteins, human-like Cav 1.1 of L-type, and Cav subunit alpha-2/delta-1 in Acanthamoeba spp. early during evolution. By in-depth investigation into genomics, transcriptomics, use of bioinformatics tools and experimentations done with drugs like amlodipine and gabapentin on Acanthamoeba spp., we show the evidence of primitive forms of these channels in this protist pathogen. Genomics and transcriptomics of proteins ACA1_167020, 092610, and 270170 reflected their cellular expression in Acanthamoeba spp. We performed amino acid sequence homology, 3D structural modeling, ligand binding predictions, and dockings. Bioinformatics and 3D structural models show similarities between ACA1_167020, 092610, 270170, and different types of known human Cav. We show amoebicidal effects of amlodipine and gabapentin on Acanthamoeba spp., which can help design their structural analogs to target pathogenic genotypes of Acanthamoeba in diseases like Acanthamoeba keratitis and granulomatous amoebic encephalitis.
真核生物电压门控钙通道（Cav）的进化是全世界生物学家感兴趣的领域。CLAN CL0030 及其家族 Ion_Trans 2 PF 07885 已知存在于原核生物中，但这些离子通道在棘阿米巴属中的起源尚未确定。我们推断了类似于蛋白质的两孔通道的原始形式、L 型的人类样 Cav 1.1 和 Cav 亚基 alpha-2/delta-1 的起源，这些在棘阿米巴属中早期进化。通过深入研究基因组学、转录组学、使用生物信息学工具以及用氨氯地平和加巴喷丁等药物对棘阿米巴属进行实验，我们证明了这些通道在这种原生动物病原体中的原始形式的证据。蛋白 ACA1_167020、092610 和 270170 的基因组学和转录组学反映了它们在棘阿米巴属中的细胞表达。我们进行了氨基酸序列同源性、3D 结构建模、配体结合预测和对接。生物信息学和 3D 结构模型显示 ACA1_167020、092610、270170 与不同类型的已知人类 Cav 之间存在相似性。我们展示了氨氯地平和加巴喷丁对棘阿米巴属的杀阿米巴作用，这有助于设计它们的结构类似物，以针对棘阿米巴角膜炎和肉芽肿性阿米巴脑炎等疾病中的致病性棘阿米巴基因型。
