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Table I.

Tim8p–Tim13 complex zinc complex


Mol zinc:mol complex

Actual (−Zn)a0:1

Actual (+Zn)a0:1

Predictedb1:1

Predictedb6:1
Zinc (sample/background, ppb)c 0.792/0.352 0.821/0.443 7.34 44.06 
Zinc (μM)d 0.007 0.006 1.12 6.72 
Protein (mg/ml)e 0.071 0.074   
Protein (μM)d 1.12 1.17   
Zinc per complex
 
 .098
 
 .081
 
1
 
6
 

Mol zinc:mol complex

Actual (−Zn)a0:1

Actual (+Zn)a0:1

Predictedb1:1

Predictedb6:1
Zinc (sample/background, ppb)c 0.792/0.352 0.821/0.443 7.34 44.06 
Zinc (μM)d 0.007 0.006 1.12 6.72 
Protein (mg/ml)e 0.071 0.074   
Protein (μM)d 1.12 1.17   
Zinc per complex
 
 .098
 
 .081
 
1
 
6
 
a

Values represent an average of three independent purifications from cells induced in the absence (−Zn) or presence of 1 mM zinc-acetate (+Zn).

b

Each twin CX3C motif (six per complex) in the Tim8p–Tim13p complex could potentially coordinate one Zn2+. The predicted levels of zinc are shown (in ppb) for hypothetical Tim8p–Tim13p complexes that bind one Zn2+ per complex or one Zn2+ per monomer (six zinc ions total). Predicted values are based on the same concentration of Tim8p–Tim13p complex (−Zn) used in actual measurements.

c

The average ICP-determined zinc concentration (ppb) is presented along with the average background level of zinc. Background levels of zinc were determined using buffer blanks from the same purification.

d

The micromolar concentrations of both protein and zinc were calculated assuming a molecular mass of 65.38 atomic mass units (amu) for zinc and 6.32 × 104 amu for the Tim8p–Tim13p complex.

e

The protein concentration of each sample was determined by quantitative amino acid analysis.

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