A luminometric method for the determination of ATP and phosphocreatine in single human skeletal muscle fibres.
A sensitive method for the analysis of ATP and phosphocreatine (PCr) in single human skeletal muscle fibres is described. Muscle tissue was freeze-dried and single fibres were dissected free with the aid of low-power microscopy. The fibres were then extracted in trichloroacetic acid and neutralized with KHCO3. The assay is based on the continuous monitoring of light produced as a result of ATP degradation in the firefly luciferase reaction. PCr is measured as the amount of ATP formed in the creatine kinase reaction. The coefficient of variation was less than 4% for both ATP and PCr determination. The amount of tissue required for the assay is approximately 0.5 microgram (dry weight). The assay showed good agreement with spectrophotometric and high-performance liquid chromatographic (HPLC) measurements made upon extracts of whole muscle tissue.
Wibom
R
R
Department of Clinical Chemistry II, Huddinge University Hospital, Sweden.
Söderlund
K
K
Lundin
A
A
Hultman
E
E
D000255:Adenosine Triphosphate / Q000032:analysis* / Q000592:standards; D005069:Evaluation Studies as Topic; D006801:Humans; D008156:Luciferases; D008163:Luminescent Measurements; D009132:Muscles / Q000737:chemistry*; D010725:Phosphocreatine / Q000032:analysis* / Q000592:standards
D010725:Phosphocreatine; D000255:Adenosine Triphosphate; D008156:Luciferases