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Table I.

Glutathione content and redox state of cytosolic rxYFP in various yeast strains





Glutathione content
 (pmol/106 cells)b



StrainRelevant genotypeSuppl.aGSHGSSGGSSG/GSHOxidized rxYFPcE′cytosold

 

 

 

 

 
%
 
%
 

 
M4745 wild type  206 ± 4 9.2 ± 0.4 10 ± 1  −289 mV 
M4860       glr1  172 ± 3 33 ± 3 19 56 ± 3  −252 mV 
M4860       glr1 GSSG 265 ± 9 302 ± 5 114 84 ± 1 ND 
M4859   glr1 met15 GSH 267 ± 6 161 ± 3 60 75 ± 2 ND 
M4859   glr1 met15 GSSG 186 ± 6 417 ± 9 225 92 ± 1 ≥ −227 mV 
M5059
 
  glrl grx1 grx2
 

 
225 ± 3
 
67 ± 1
 
30
 
ND
 
−245 mV
 




Glutathione content
 (pmol/106 cells)b



StrainRelevant genotypeSuppl.aGSHGSSGGSSG/GSHOxidized rxYFPcE′cytosold

 

 

 

 

 
%
 
%
 

 
M4745 wild type  206 ± 4 9.2 ± 0.4 10 ± 1  −289 mV 
M4860       glr1  172 ± 3 33 ± 3 19 56 ± 3  −252 mV 
M4860       glr1 GSSG 265 ± 9 302 ± 5 114 84 ± 1 ND 
M4859   glr1 met15 GSH 267 ± 6 161 ± 3 60 75 ± 2 ND 
M4859   glr1 met15 GSSG 186 ± 6 417 ± 9 225 92 ± 1 ≥ −227 mV 
M5059
 
  glrl grx1 grx2
 

 
225 ± 3
 
67 ± 1
 
30
 
ND
 
−245 mV
 
a

Strains were grown in SC medium at 30°C. GSH and GSSG indicate supplementation of SC-Met medium with either 1 mM GSH or GSSG.

b

In order to calculate the amount of GSH or GSSG per 106 cells, one OD600 U was determined for M4745 to be 1.5·107 cells/ml. Values are given as the mean ± SD of measurements on three independent cultures.

c

The proportion of oxidized cytosolic rxYFP (mean ± SD; n = 3) was determined by spectrofluorometric redox titration as described in Materials and methods.

d

Cytosolic redox potentials (E′cytosol) were calculated from the pulse-chase end point redox distributions given in Figs. 2 and 3 and an E°′ of −265 mV for rxYFP (Table II).

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