Document Detail

A re-evaluation of conditions required for an accurate estimation of the extramitochondrial ATP/ADP ratio in isolated rat-liver mitochondria.
MedLine Citation:
PMID:  6091749     Owner:  NLM     Status:  MEDLINE    
The values reported in the literature for the extramitochondrial ATP/ADP ratio in resting rat-liver mitochondria (State 4) vary widely. The conditions required for an accurate determination of this parameter were therefore investigated. In experiments with rat-liver mitochondria incubated under State-4 conditions, it was found that the extramitochondrial ATP/ADP ratio, as calculated from the values measured in neutralised perchloric acid extracts, was lower than that estimated from the concentrations of creatine and creatine phosphate, using the metabolite indicator method. The discrepancy is due to hydrolysis of ATP occurring in the presence of perchloric acid. Conditions are described for minimising ATP hydrolysis in the presence of perchloric acid, and include the use of low concentrations of perchloric acid, short times of exposure to the acid before neutralisation, low temperatures and the presence of excess EDTA. Under these conditions, the values obtained for the extramitochondrial ATP/ADP ratio agreed with those calculated by the metabolite indicator method, provided ratios do not exceed the value of 100. In cases where the extramitochondrial ATP/ADP does exceed 100, phenol/chloroform/isoamyl alcohol must be used to quench the reactions, as described by Slater et al. (Slater, E.C., Rosing, J. and Mol, A. (1973) Biochim. Biophys. Acta 292, 534-553). With this method, the extramitochondrial ATP/ADP ratio was found to have a value of more than 1000 in rat-liver mitochondria incubated with succinate + rotenone in the resting state (pH 7.0; T = 37 degrees C), in agreement with Slater et al.
R J Wanders; G B Van den Berg; J M Tager
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biochimica et biophysica acta     Volume:  767     ISSN:  0006-3002     ISO Abbreviation:  Biochim. Biophys. Acta     Publication Date:  1984 Oct 
Date Detail:
Created Date:  1984-12-06     Completed Date:  1984-12-06     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0217513     Medline TA:  Biochim Biophys Acta     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  113-9     Citation Subset:  IM    
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MeSH Terms
Adenosine Diphosphate / metabolism*
Adenosine Triphosphate / metabolism*
Cold Temperature
Creatine / metabolism
Edetic Acid
False Negative Reactions
Mitochondria, Liver / metabolism*
Perchloric Acid
Phosphocreatine / metabolism
Rats, Inbred Strains
Reg. No./Substance:
56-65-5/Adenosine Triphosphate; 57-00-1/Creatine; 58-64-0/Adenosine Diphosphate; 60-00-4/Edetic Acid; 67-07-2/Phosphocreatine; 7601-90-3/Perchloric Acid

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