Low apparent creatine kinase activity and prolonged lag phases in serum of patients with metastatic disease: elimination by treatment of sera with sulfhydryl agents.
We find lag phases exceeding 20 min in measuring creatine kinase activity, by using the kinetic creatin phosphate leads to creatine assay, in sera of some patients with carcinoma metastatic to the liver. Such long lag phases are accompanied by a decreased apparent enzyme activity. These problems are eliminated by adding sulfhydryl agents to the serum before assay, but not by adding more of such agents to the assay reagent. beta-Mercaptoethanol is supperior to Cleland's reagents, glutathione, and cysteine. The long lag phases could not be explained by inadequate activity of the coupling enzymes, interference with the coupling steps, high proportions of cardiacisoenzymes activity, simple oxidation of the enzyme, low concentrations of albumin, or increased concentrations of glutathione reductase, lactate dehydrogenase, or uric acid. We conclude that the prolonged lag phases reflect inadequate reactivation of the enzyme by sulfhydryl agents under the usual assay conditions. Reactivation before assay can prevent potentially serious negative errors in the assay of creatine kinase.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D003402:Creatine Kinase / Q000097:blood* / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D005188:False Negative Reactions; D005956:Glucose-6-Phosphate Isomerase / Q000097:blood; D006801:Humans; D007700:Kinetics; D008623:Mercaptoethanol / Q000494:pharmacology; D009362:Neoplasm Metastasis / Q000201:enzymology*; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D013997:Time Factors
D013438:Sulfhydryl Compounds; D008623:Mercaptoethanol; D000255:Adenosine Triphosphate; D003402:Creatine Kinase; D005956:Glucose-6-Phosphate Isomerase; D004229:Dithiothreitol