Tags

Type your tag names separated by a space and hit enter

Evaluation of methods for the determination of mitochondrial respiratory chain enzyme activities in human skeletal muscle samples.
Anal Biochem. 2000 Mar 01; 279(1):55-60.AB

Abstract

The quantification of mitochondrial enzyme activities in skeletal muscle samples of patients suspected of having mitochondrial myopathies is problematic. Therefore, we have evaluated different methods for the determination of activities cytochrome c oxidase and NADH:CoQ oxidoreductase in human skeletal muscle samples. The measurement of cytochrome c oxidase activity in the presence of 200 microM ferrocytochrome c and the detection of NADH:CoQ oxidoreductase as rotenone-sensitive NADH:CoQ(1) reductase resulted in comparable citrate synthase-normalized respiratory chain enzyme activities of both isolated mitochondria and homogenates from control human skeletal muscle samples. These methods allowed the precise detection of deficiencies of respiratory chain enzymes in skeletal muscle of two patients harboring only 20 and 27% of deleted mitochondrial DNA, respectively. Therefore, citrate synthase-normalized respiratory chain activities can serve as stable reference values for the determination of a putative mitochondrial defect in human skeletal muscle.

Authors+Show Affiliations

Department of Epileptology, Department of Neurology, University Bonn Medical Center, Sigmund-Freud-Strasse 25, Bonn, D-53105, Germany.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10683230

Citation

Wiedemann, F R., et al. "Evaluation of Methods for the Determination of Mitochondrial Respiratory Chain Enzyme Activities in Human Skeletal Muscle Samples." Analytical Biochemistry, vol. 279, no. 1, 2000, pp. 55-60.
Wiedemann FR, Vielhaber S, Schröder R, et al. Evaluation of methods for the determination of mitochondrial respiratory chain enzyme activities in human skeletal muscle samples. Anal Biochem. 2000;279(1):55-60.
Wiedemann, F. R., Vielhaber, S., Schröder, R., Elger, C. E., & Kunz, W. S. (2000). Evaluation of methods for the determination of mitochondrial respiratory chain enzyme activities in human skeletal muscle samples. Analytical Biochemistry, 279(1), 55-60.
Wiedemann FR, et al. Evaluation of Methods for the Determination of Mitochondrial Respiratory Chain Enzyme Activities in Human Skeletal Muscle Samples. Anal Biochem. 2000 Mar 1;279(1):55-60. PubMed PMID: 10683230.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Evaluation of methods for the determination of mitochondrial respiratory chain enzyme activities in human skeletal muscle samples. AU - Wiedemann,F R, AU - Vielhaber,S, AU - Schröder,R, AU - Elger,C E, AU - Kunz,W S, PY - 2000/2/23/pubmed PY - 2000/5/16/medline PY - 2000/2/23/entrez SP - 55 EP - 60 JF - Analytical biochemistry JO - Anal Biochem VL - 279 IS - 1 N2 - The quantification of mitochondrial enzyme activities in skeletal muscle samples of patients suspected of having mitochondrial myopathies is problematic. Therefore, we have evaluated different methods for the determination of activities cytochrome c oxidase and NADH:CoQ oxidoreductase in human skeletal muscle samples. The measurement of cytochrome c oxidase activity in the presence of 200 microM ferrocytochrome c and the detection of NADH:CoQ oxidoreductase as rotenone-sensitive NADH:CoQ(1) reductase resulted in comparable citrate synthase-normalized respiratory chain enzyme activities of both isolated mitochondria and homogenates from control human skeletal muscle samples. These methods allowed the precise detection of deficiencies of respiratory chain enzymes in skeletal muscle of two patients harboring only 20 and 27% of deleted mitochondrial DNA, respectively. Therefore, citrate synthase-normalized respiratory chain activities can serve as stable reference values for the determination of a putative mitochondrial defect in human skeletal muscle. SN - 0003-2697 UR - https://www.unboundmedicine.com/medline/citation/10683230/Evaluation_of_methods_for_the_determination_of_mitochondrial_respiratory_chain_enzyme_activities_in_human_skeletal_muscle_samples_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-2697(99)94434-2 DB - PRIME DP - Unbound Medicine ER -