Gragg Megan, Park Paul S-H
Department of Ophthalmology and Visual Sciences, Case Western Reserve University, Cleveland, OH, United States.
Department of Ophthalmology and Visual Sciences, Case Western Reserve University, Cleveland, OH, United States.
Methods Cell Biol. 2019;149:87-105. doi: 10.1016/bs.mcb.2018.08.007. Epub 2018 Sep 17.
Rhodopsin is the light receptor in rod photoreceptor cells of the retina that plays a central role in phototransduction and rod photoreceptor cell health. Rhodopsin mutations are the leading known cause of autosomal dominant retinitis pigmentosa, a retinal degenerative disease. A majority of rhodopsin mutations cause misfolding and aggregation of the apoprotein opsin. The nature of aggregates formed by misfolded rhodopsin mutants and the associated cell toxicity is poorly understood. Misfolding rhodopsin mutants have been characterized biochemically, and categorized as either partial or complete misfolding mutants. This classification is incomplete and does not provide sufficient information to fully understand rhodopsin aggregation, disease pathogenesis, and evaluate therapeutic strategies. To better understand the aggregation of misfolded rhodopsin mutants, a Förster resonance energy transfer assay has been developed to monitor the aggregation of fluorescently tagged mutant rhodopsins expressed in live cells.
视紫红质是视网膜视杆光感受器细胞中的光受体，在光转导和视杆光感受器细胞健康中起核心作用。视紫红质突变是常染色体显性视网膜色素变性（一种视网膜退行性疾病）的主要已知病因。大多数视紫红质突变会导致脱辅基蛋白视蛋白错误折叠和聚集。由错误折叠的视紫红质突变体形成的聚集体的性质以及相关的细胞毒性尚不清楚。错误折叠的视紫红质突变体已通过生物化学方法进行了表征，并被分类为部分或完全错误折叠突变体。这种分类是不完整的，并且没有提供足够的信息来充分理解视紫红质聚集、疾病发病机制以及评估治疗策略。为了更好地理解错误折叠的视紫红质突变体的聚集情况，已经开发了一种Förster共振能量转移测定法来监测在活细胞中表达的荧光标记突变视紫红质的聚集。
