Table 1.
Purification of lysosomes from cultured ARPE-19 cells
| Marker | Marker for | Sample | Homogenate (specific activity) | Lysosomal Fraction (specific activity) |
|---|---|---|---|---|
| β-Hexosaminidase | Lysosomes | Control / 24h | 2.4 mU/mg protein | 261.2 mU/mg protein |
| EC 3.2.1.30 | Control / 48h | 2.5 mU/mg protein | 273.5 mU/mg protein | |
| MG-132 / 24h | 2.4 mU/mg protein | 243.2 mU/mg protein | ||
| MG-132 / 48 | 2.3 mU/mg protein | 241.4 mU/mg protein | ||
| Succinate dehydrogenase | Mitochondria | Control / 24h | 0.75 mU/mg protein | 0.02 mU/mg protein |
| E.C. 1.3.99.1 | Control / 48h | 0.71 mU/mg protein | 0.04 mU/mg protein | |
| MG-132 / 24h | 0.68 mU/mg protein | 0.02 mU/mg protein | ||
| MG-132 / 48 | 0.65 mU/mg protein | 0.03 mU/mg protein | ||
| Lactate dehydrogenase | Cytosol | Control / 24h | 2078 mU/mg protein | n.d. |
| EC 1.1.1.27 | Control / 48h | 1967 mU/mg protein | n.d. | |
| MG-132 / 24h | 1898 mU/mg protein | n.d. | ||
| MG-132 / 48 | 1912 mU/mg protein | n.d. | ||
| Alkaline phosphodiesterase | Plasma membrane | Control / 24h | 628 μU/mg protein | n.d. |
| EC 3.1.4.1 | Control / 48h | 598 μU/mg protein | n.d. | |
| MG-132 / 24h | 605 μU/mg protein | n.d. | ||
| MG-132 / 48 | 611 μU/mg protein | n.d. | ||
| UDP-galactosyltransferase | Microsomes (ER/Golgi) | Control / 24h | 69 nU/mg protein | n.d. |
| E.C. 2.4.1.38 | Control / 48h | 61 nU/mg protein | n.d. | |
| MG-132 / 24h | 65 nU/mg protein | n.d. | ||
| MG-132 / 48 | 64 nU/mg protein | n.d. | ||
| DNA | Nuclei | Control / 24h | 3.48 μg/mg protein | n.d. |
| Control / 48h | 3.46 μg/mg protein | n.d. | ||
| MG-132 / 24h | 3.49 μg/mg protein | n.d. | ||
| MG-132 / 48 | 3.48 μg/mg protein | n.d. |
n.d. not detectable.