Ashmus Roger A, Shen David L, Vocadlo David J
Simon Fraser University, Burnaby, BC, Canada.
Simon Fraser University, Burnaby, BC, Canada.
Methods Enzymol. 2018;598:199-215. doi: 10.1016/bs.mie.2017.06.014. Epub 2017 Oct 31.
Glucocerebrosidase (GCase) is a lysosomal glycoside hydrolase that cleaves the glycolipid glucosylceramide (GlcCer). Deficiencies of this enzyme lead to accumulation of GlcCer and the development of the lysosomal storage disease known as Gaucher's disease. Recently, loss-of-function mutations in the GBA1 gene that encodes GCase have been linked to Parkinson's disease. Currently pursued therapeutic strategies to increase GCase involve enzyme replacement therapy, chemical chaperone therapy, and GCase activators. A challenge associated with advancing such strategies is to efficiently monitor GCase activity within the lysosomes of live cells. In this chapter, we review the design and use of the fluorescent-quenched probe GBA1-FQ2 to quantitatively measure GCase activity in lysosomes of live cells.
葡糖脑苷脂酶（GCase）是一种溶酶体糖苷水解酶，可裂解糖脂葡萄糖神经酰胺（GlcCer）。该酶的缺乏会导致GlcCer积累，并引发溶酶体贮积病，即戈谢病。最近，编码GCase的GBA1基因功能丧失突变与帕金森病有关。目前正在探索的增加GCase的治疗策略包括酶替代疗法、化学伴侣疗法和GCase激活剂。推进这些策略面临的一个挑战是有效监测活细胞溶酶体内的GCase活性。在本章中，我们将回顾荧光淬灭探针GBA1-FQ2的设计和用途，以定量测量活细胞溶酶体内的GCase活性。
