Kurnia Ryan Septa, Soebandrio Amin, Harun Vivi Hardianty, Nugroho Christian Marco Hadi, Krisnamurti Desak Gede Budi, Poetri Okti Nadia, Indrawati Agustin, Tarigan Simson, Natih Ketut Karuni Nyanakumari, Ibrahim Fera, Sudarmono Pratiwi Pudjilestari, Silaen Otto Sahat Martua
Doctoral Program in Biomedical Science, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia.
Department of Microbiology, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia.
J Adv Vet Anim Res. 2023 Dec 31;10(4):667-676. doi: 10.5455/javar.2023.j722. eCollection 2023 Dec.
This study aims to evaluate the effect of sialidase treatment on monolayer cell behavior using computational screening and an approach to demonstrate interaction between enzyme-based drugs and ligands in host cells.
The study was carried out by molecular docking analysis used to predict the interactions between atoms that occur, followed by genetic characterization of sialidase from a wild isolate. Sialidase, which has undergone further production and purification processes exposed to chicken embryonic fibroblast cell culture, and observations-based structural morphology of cells compared between treated cells and normal cells without treatment.
Based on an study, sialidase has an excellent binding affinity with Neu5Acα (2.3) Gal ligand receptor with Gibbs energy value (∆G)-7.35 kcal/mol and Ki value of 4.11 µM. Wild isolates in this study have 99.1%-100% similarity to the plc gene, NanH, and NanI genes, while NanJ shows 93.18% similarity compared to the reference isolate from GenBank. Sialidase at 750 and 150 mU may impact the viability, cell count, and cell behavior structure of fibroblast cells by significantly increasing the empty area and perimeter of chicken embryo fibroblast (CEF) cells, while at 30 mU sialidase shows no significant difference compared with mock control.
Sialidase-derived has the capacity to compete with viral molecules for attachment to host sialic acid based on analysis. However, sialidase treatment has an impact on monolayer cell fibroblasts given exposure to high doses.
本研究旨在通过计算筛选以及一种用于证明基于酶的药物与宿主细胞中配体之间相互作用的方法，评估唾液酸酶处理对单层细胞行为的影响。
该研究通过分子对接分析来预测原子间发生的相互作用，随后对野生分离株的唾液酸酶进行基因表征。经过进一步生产和纯化过程的唾液酸酶暴露于鸡胚成纤维细胞培养中，并比较处理细胞与未处理的正常细胞之间基于观察的细胞结构形态。
基于一项研究，唾液酸酶与Neu5Acα(2.3)Gal配体受体具有出色的结合亲和力，吉布斯自由能值(∆G)为 -7.35 kcal/mol，Ki值为4.11 µM。本研究中的野生分离株与plc基因、NanH和NanI基因具有99.1%-100%的相似性，而与来自GenBank的参考分离株相比，NanJ显示出93.18%的相似性。750和150 mU的唾液酸酶可能会通过显著增加鸡胚成纤维细胞(CEF)的空泡面积和周长来影响成纤维细胞的活力、细胞数量和细胞行为结构，而30 mU的唾液酸酶与模拟对照相比无显著差异。
基于分析，源自唾液酸酶的物质有能力与病毒分子竞争附着于宿主唾液酸。然而，唾液酸酶处理在高剂量暴露时会对单层细胞成纤维细胞产生影响。
