Sharma Ritika, Kumari Anchala, Kundu Bishwajit, Grover Abhinav
School of Biotechnology, Jawaharlal Nehru University New Delhi-110067 India
Indian Council of Medical Research, International Health Division New Delhi-110029 India
RSC Adv. 2022 Oct 14;12(45):29469-29481. doi: 10.1039/d2ra05061g. eCollection 2022 Oct 11.
Inherited glaucoma is a recent addition to the inventory of diseases arising due to protein misfolding. Mutations in the olfactomedin (OLF) domain of myocilin are the most common genetic cause behind this disease. Disease associated variants of m-OLF are predisposed to misfold and aggregate in the trabecular meshwork (TM) tissue of the eye. In recent years, the nature of these aggregates was revealed to exhibit the hallmarks of amyloids. Amyloid aggregates are highly stable structures that are formed, often with toxic consequences in a number of debilitating diseases. In spite of its clinical relevance the amyloidogenic nature of m-OLF has not been studied adequately. Here we have studied the amyloid fibrillation of m-OLF and report ECG as an inhibitor against it. Using biophysical and biochemical assays, coupled with advanced microscopic evaluations we show that ECG binds and stabilizes native m-OLF and thus prevents its aggregation into amyloid fibrils. Furthermore, we have used REMD simulations to delineate the stabilizing effects of ECG on the structure of m-OLF. Collectively, we report ECG as a molecular scaffold for designing and testing of novel inhibitors against m-OLF amyloid fibrillation.
遗传性青光眼是蛋白质错误折叠引发的疾病清单中的新增成员。肌纤蛋白(MYOC)嗅觉介质(OLF)结构域的突变是该疾病最常见的遗传病因。与疾病相关的m-OLF变体易于在眼部小梁网(TM)组织中错误折叠并聚集。近年来,这些聚集体的性质被揭示具有淀粉样蛋白的特征。淀粉样聚集体是高度稳定的结构,其形成往往会在许多使人衰弱的疾病中产生毒性后果。尽管m-OLF的淀粉样变性性质具有临床相关性,但尚未得到充分研究。在此,我们研究了m-OLF的淀粉样纤维化,并报告了依西美坦作为其抑制剂。通过生物物理和生化分析,结合先进的显微镜评估,我们表明依西美坦结合并稳定天然m-OLF,从而防止其聚集形成淀粉样纤维。此外,我们使用了复制交换分子动力学(REMD)模拟来描述依西美坦对m-OLF结构的稳定作用。我们共同报告依西美坦作为一种分子支架,用于设计和测试针对m-OLF淀粉样纤维化的新型抑制剂。