Blyden K'Mani, Li Susanna, Gomez Claudia, Jain Samyak, Bhattacharya Sanjoy K
Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
Medical College of Georgia, Augusta University, Augusta, Georgia.
Methods Mol Biol. 2025;2925:103-112. doi: 10.1007/978-1-0716-4534-5_6.
Pseudoexfoliation syndrome (PEX) is an age-related ocular disorder marked by the formation of proteinaceous extracellular deposits on the anterior ocular structures and other organs. Although the exact pathogenic mechanism of PEX deposit formation remains unclear, these deposits can elevate intraocular pressure, potentially leading to optic nerve degeneration and pseudoexfoliation glaucoma (PEXG), a severe form of open-angle glaucoma. Essential proteins implicated in PEX include lysyl oxidase-like 1, clusterin, and fibulin-5. PEX also possesses a unique metabolome, making metabolite analysis crucial to understanding the biochemical changes and pathways involved in its pathogenesis. Recent theories suggest that aberrant vitamin-D-binding protein may act as a nucleation center for protein clumping, exacerbating PEX. Advanced techniques like liquid chromatography-mass spectrometry are vital for profiling the metabolomic, proteomic, and lipidomic composition of PEX and PEXG deposits, providing insights into disease mechanisms and potential treatments. We aim to review methods for metabolite and protein extraction in addition to metabolic and proteomic analysis utilized in distinguishing the compositional differences between PEX and PEXG deposits and to understand the transition between these conditions. Such metabolomic analyses may play a crucial role in identifying potential biomarkers and potential therapeutic targets. In this book chapter, we discuss the methods to characterize extraction of metabolite associated with protein deposits and their analysis.
假性剥脱综合征(PEX)是一种与年龄相关的眼部疾病,其特征是在前眼部结构和其他器官上形成蛋白质细胞外沉积物。尽管PEX沉积物形成的确切致病机制尚不清楚,但这些沉积物可升高眼压,可能导致视神经变性和假性剥脱性青光眼(PEXG),这是开角型青光眼的一种严重形式。与PEX相关的重要蛋白质包括赖氨酰氧化酶样1、簇集蛋白和纤连蛋白-5。PEX还具有独特的代谢组,因此代谢物分析对于理解其发病机制中涉及的生化变化和途径至关重要。最近的理论表明,异常的维生素D结合蛋白可能作为蛋白质聚集的成核中心,加剧PEX。液相色谱-质谱等先进技术对于分析PEX和PEXG沉积物的代谢组学、蛋白质组学和脂质组学组成至关重要,有助于深入了解疾病机制和潜在治疗方法。我们旨在回顾除了用于区分PEX和PEXG沉积物组成差异的代谢和蛋白质组分析之外的代谢物和蛋白质提取方法,并了解这些病症之间的转变。这种代谢组学分析可能在识别潜在生物标志物和潜在治疗靶点方面发挥关键作用。在本章中,我们讨论了表征与蛋白质沉积物相关的代谢物提取及其分析的方法。