Table 1.
Isolation and enrichment of RNase P
| Fraction
|
Volume (ml)
|
Total units (103)a
|
Total protein (mg)
|
Sp. Act. (U/mg)
|
Enrichment (fold)
|
|---|---|---|---|---|---|
| Yeast extract | 498 | 41.1 | 23,700 | 1.74 | 1 |
| SP—Sepharose | 775 | 53.9 | 852 | 63.3 | 36 |
| DE52 | 94 | 16.4 | 167 | 97.9 | 56 |
| Resource Q | 10.5 | 8.1 | 11.7 | 693 | 400 |
| Resource S | 2.9 | 2.3 | 0.3 | 6760 | 3900 |
| Glycerol gradient peak | 16 | 0.65 | [0.001]b | [650,000]b | [370,000]b |
a
1 Unit = relative RNase P activity required to convert 10% of 1.0 pmole of pre-tRNAASP in 15 min under standard assay conditions.
b
Quantity of total protein in glycerol gradient peak estimated on the basis of total RNA (see Materials and Methods).