Table 1. Summary of Fe Percentages and Concentrations Determined by Biophysical Methodsa.
| sample | total [Fe] (ICP-MS) | NHHS FeII (MB) | HS FeII hemes (MB) | CD (MB) | nanoparticles (MB) | NHHS FeIII (MB) | FeII hemes (UV–vis) |
g = 4.3 (EPR) |
|---|---|---|---|---|---|---|---|---|
| WT1 | 170 ± 45 μM (6) | 25% (43 μM) | 5% (9 μM) | 60% (103 μM) | <5% (<9 μM) | 0% | 44 ± 1 μM | 0 μM |
| WT40 | 490 ± 140 μM (8) | 13% (64 μM) | 3% (15 μM) | 8% (39 μM) | 18% (88 μM) | 55% (269 μM) | 55 ± 3 μM | 152 μM |
| Δ1 | 148 ± 64 μM (8) | 15% (22 μM) | 10% (15 μM) | 57% (85 μM) | <5% (<7 μM) | 0% | 46 ± 2 μM | <1 μM |
| Δ10 | 209 ± 8 μM (2) | 26% (54 μM) | 7% (15 μM) | 39% (82 μM) | 5% (11 μM) | 23% (48 μM) | ND | ND |
| Δ20 | 285 ± 5 μM (2) | 35% (100 μM) | 5% (14 μM) | 27% (77 μM) | 8% (23 μM) | 28% (80 μM) | ND | ND |
| Δ40 | 360 ± 100 μM (7) | 30% (107 μM) | 5% (18 μM) | 19% (69 μM) | 17% (61 μM) | 30% (108 μM) | 43 ± 4 μM | 73 μM |
| Δ40, Mitos | 854 ± 102 μM (2) | 37% (316 μM) | 5% (43 μM) | 32% (273 μM) | 23% (197 μM) | 0% | 364 ± 9 μM | ND |
| UP1 | 250 ± 60 μM (3) | 51% (130 μM) | 3% (10 μM) | 10% (30 μM) | 19% (50 μM) | 18% (50 μM) | 32 ± 3 μM | 30 μM |
| UP10 | 565 ± 10 μM (2) | 53% (300 μM) | 2% (11 μM) | 5% (28 μM) | 25% (141 μM) | 18% (102 μM) | ND | ND |
| UP20 | 979 ± 24 μM (2) | 39% (382 μM) | ≤1% (≤10 μM) | 3% (29 μM) | 39% (382 μM) | 17% (167 μM) | ND | ND |
| UP40 | 1839 ± 750 μM (5) | 38% (700 μM) | ≤1% (≤18 μM) | 2% (37 μM) | 24% (441 μM) | 17% (313 μM) | 31 ± 6 μM | 372 μM |
| UP1+A | 385 ± 68 μM (3) | 70% (270 μM) | 2% (8 μM) | 7% (27 μM) | 11% (42 μM) | 10% (39 μM) | 29 ± 3 μM | 39 μM |
| UP40+A | 1040 ± 108 μM (4) | 18% (187 μM) | 2% (21 μM) | 4% (42 μM) | 29% (301 μM) | 45% (468 μM) | 40 ± 4 μM | 415 μM |
a
In the column “total [Fe]”, the number of replicates are indicated in parentheses. Uncertainties in Mössbauer percentages and corresponding concentrations (in parentheses) are ±3%. Concentrations of FeII hemes, as determined by UV-vis, are the sum of individual components (hemes a, b, and c), simulated as described.24 The g = 4.3 EPR signals were quantified from individual samples (Table S3) as described.24 Sample concentrations were corrected for packing efficiencies: 0.73 for whole cells, 0.75 for isolated vacuoles, and 0.78 for isolated mitochondria.9,23,26 ND = not determined.