Environmental and Life Sciences Graduate Program, Trent University, Peterborough, ON, Canada.
Department of Biology, Trent University, Peterborough, ON, Canada.
Methods Mol Biol. 2024;2814:55-79. doi: 10.1007/978-1-0716-3894-1_4.
Lysosomes are membrane-enclosed organelles that digest intracellular material. They contain more than 50 different enzymes that can degrade a variety of macromolecules including nucleic acids, proteins, polysaccharides, and lipids. In addition to functioning within lysosomes, lysosomal enzymes are also secreted. Alterations in the levels and activities of lysosomal enzymes dysregulates lysosomes, which can lead to the intralysosomal accumulation of biological material and the development of lysosomal storage diseases (LSDs) in humans. Dictyostelium discoideum has a long history of being used to study the trafficking and functions of lysosomal enzymes. More recently, it has been used as a model system to study several LSDs. In this chapter, we outline the methods for assessing the activity of several lysosomal enzymes in D. discoideum (α-galactosidase, β-galactosidase, α-glucosidase, β-glucosidase, β-N-acetylglucosaminidase, α-mannosidase, cathepsin B, cathepsin D, cathepsin F, palmitoyl protein thioesterase 1, and tripeptidyl peptidase 1).
溶酶体是一种膜结合的细胞器，可消化细胞内物质。它们包含 50 多种不同的酶，可以降解包括核酸、蛋白质、多糖和脂类在内的多种大分子。除了在溶酶体内发挥作用外，溶酶体酶也会被分泌。溶酶体酶的水平和活性的改变会导致溶酶体功能失调，这可能导致细胞内生物物质的积累，并导致人类溶酶体贮积症（LSD）的发生。盘基网柄菌（Dictyostelium discoideum）长期以来一直被用于研究溶酶体酶的运输和功能。最近，它被用作研究几种 LSD 的模型系统。在本章中，我们概述了评估盘基网柄菌中几种溶酶体酶活性的方法（α-半乳糖苷酶、β-半乳糖苷酶、α-葡萄糖苷酶、β-葡萄糖苷酶、β-N-乙酰氨基葡萄糖苷酶、α-甘露糖苷酶、组织蛋白酶 B、组织蛋白酶 D、组织蛋白酶 F、棕榈酰蛋白硫酯酶 1 和三肽基肽酶 1）。
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