TABLE 2.
Quality control of yeast subcellular fractions
For growth conditions and the isolation of subcellular compartments, see “Experimental Procedures.” The relative enrichment of markers in the homogenate was set as 1. Marker proteins were as follows: Gas1p (β-1,3-glucanosyltransferase (marker for plasma membrane)); Wbp1p (β-subunit of the oligosaccharyl transferase glycoprotein complex (marker for microsomes)); Por1p (porin (marker for mitochondria)); Erg6p (Δ(24)-sterol C-methyltransferase (marker for lipid particles)). ND, not detectable.
| Relative enrichment |
||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Wild type |
hem1Δ |
QM |
QMhem1Δ |
|||||||||||
| Gas1p | Wbp1p | Por1p | Erg6p | Gas1p | Wbp1p | Por1p | Erg6p | Gas1p | Wbp1p | Por1p | Gas1p | Wbp1p | Por1p | |
| -fold | -fold | -fold | -fold | |||||||||||
| Mitochondria | ND | 1.0 ± 0.66 | 3.8 ± 1.5 | ND | ND | 1.0 ± 0.40 | 3.2 ± 0.91 | 0.6 | ND | 1.5 ± 0.75 | 2.3 ± 0.43 | 2.0 | 2.1 ± 0.043 | 2.0 ± 0.50 |
| Microsomes, 30.000 × g | ND | 2.5 ± 0.38 | 0.5 ± 0.2 | ND | ND | 1.3 ± 0.24 | 0.1 ± 0.09 | 0.4 | ND | 3.0 ± 0.40 | ND | 1.0 | 2.9 ± 0.96 | 1.2 ± 0.20 |
| Microsomes, 40.000 × g | ND | 1.9 ± 0.075 | ND | ND | ND | 1.1 ± 0.24 | ND | ND | ND | 2.8 ± 0.96 | ND | ND | 1.1 ± 0.53 | 0.2 ± 0.058 |
| Microsomes, 100.000 × g | ND | 0.7 ± 0.001 | ND | ND | ND | 0.3 ± 0.2 | ND | ND | ND | ND | ND | ND | ND | 0.1 ± 0.08 |
| Cytosol | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND |
| Plasma membrane | ∼100 | 1.2 ± 0.14 | 0.3 ± 0.07 | ND | ∼100 | 2.5 ± 1.1 | 2.7 ± 1.2 | ND | ∼100 | 2.3 ± 1.1 | 2.5 ± 0.78 | ∼100 | 1.1 ± 0.66 | 1.2 ± 0.75 |
| Lipid particles | ND | 1.5 ± 0.75 | 0.5 ± 0.3 | ∼100 | ND | 1.0 ± 0.60 | 0.5 ± 0.3 | ∼100 | ||||||