Table 3.
Percentage of plasma acid hydrolases that specifically bind to a CI-MPR affinity column (1 representative experiment, n = 3
| A. Effect of endoglycosydase H, alkaline phosphatase (Alk.phos.), and uncovering enzyme treatment | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Wild-type |
UCE −/− |
|||||||||
| Untreated | Endo Ha | Alk.phos. | UCE | UCE +Alk.phos. | Untreated | Endo Ha | Alk.phos. | UCE | UCE +Alk.phos. | |
| β-Mannosidase | 50 | 0 | 12 | 41 | 2 | 9 | 0 | 9 | 71 | 14 |
| β-Glucuronidase | 49 | 0 | 5 | 47 | 8 | 79 | 0 | 77 | 88 | 34 |
| β-Hexosaminidase | 12 | 3 | 4 | 8 | 1 | 12 | 1 | 3 | 36 | 3 |
| B. Inhibition of binding by Man-6-P (M6P) or GlcNAc-P-α-methylmannose | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Wild-type |
UCE −/− |
|||||||||
| Untreated | 200 μMM6P | 10 mMGlcNAc-P-α-methylman | 1 mMM6P | Untreated | 200 μMM6P | 10 mMGlcNAc-P-α-methylman | 1 mMM6P | 200 μM M6P + 10mM GlcNAc-P-α-methylman | ||
| β-Mannosidase | 50 | 10 | 49 | 0 | 9 | 14 | 1 | 1 | 1 | |
| β-Glucuronidase | 49 | 17 | 50 | 11 | 79 | 82 | 50 | 65 | 28 | |
| β-Hexosaminidase | 12 | 0 | 14 | 3 | 12 | 6 | 5 | 4 | 3 | |
a n = 2.