Document Detail

Dependence of mitochondrial and cytosolic adenine nucleotides on oxygen partial pressure in isolated hepatocytes. Application of a new rapid high pressure filtration technique for fractionation.
MedLine Citation:
PMID:  3390136     Owner:  NLM     Status:  MEDLINE    
By using a new rapid high pressure filtration technique, mitochondrial and cytosolic ATP and ADP contents were determined in isolated hepatocytes at different oxygen partial pressures. At 670 mmHg, subcellular adenine nucleotide contents and ATP/ADP ratios were comparable with values obtained with the digitonin fractionation technique. However at lower oxygen partial pressure ADP appears to be rephosphorylated during digitonin fractionation whereas with high pressure filtration fractionation rephosphorylation of ADP is avoided due to shorter fractionation times. Cytosolic and mitochondrial ATP/ADP ratios decrease if oxygen partial pressure is lowered. However the absolute values of ATP/ADP ratios depend critically on the incubation conditions. Thus incubation of hepatocytes in an oxystat system, where oxygen partial pressure is maintained constant by infusing oxygen-saturated medium and the hepatocyte suspension is continuously stirred, yields much higher subcellular and overall ATP/ADP ratios than incubation in Erlenmeyer flasks gassed with different gas mixtures and shaken in a water bath. This is ascribed to limited diffusion of oxygen from the medium into the cell if the suspension is not mixed thoroughly by stirring. The strong dependence of subcellular ATP/ADP ratios on incubation conditions indicates that oxygen may be one rate-controlling factor for oxidative phosphorylation in the intact cell.
H Hummerich; H de Groot; T Noll; S Soboll
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Biochemical journal     Volume:  250     ISSN:  0264-6021     ISO Abbreviation:  Biochem. J.     Publication Date:  1988 Mar 
Date Detail:
Created Date:  1988-08-05     Completed Date:  1988-08-05     Revised Date:  2010-09-10    
Medline Journal Info:
Nlm Unique ID:  2984726R     Medline TA:  Biochem J     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  641-5     Citation Subset:  IM    
Institut für Physiologische Chemie I der Universität Düsseldorf, Federal Republic of Germany.
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MeSH Terms
Adenine Nucleotides / metabolism*
Adenosine Triphosphate / metabolism
Cell Fractionation / methods
Cytosol / metabolism
Filtration / methods
Liver / metabolism*
Mitochondria, Liver / metabolism
Oxygen / metabolism*
Partial Pressure
Rats, Inbred Strains
Reg. No./Substance:
0/Adenine Nucleotides; 11024-24-1/Digitonin; 56-65-5/Adenosine Triphosphate; 7782-44-7/Oxygen

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