Abstract
The pyruvate dehydrogenase complex plays a central role in the pyruvate oxidation. The complex consists of five components of which pyruvate dehydrogenase (E1) catalyzes the rate limiting step.
Since E1 has a low activity in human skeletal muscle tissue a sensitive method is needed for diagnostic purposes.
Measurement of 14CO2-production from [1-14C]pyruvate provides a specific and sensitive assay for measuring E1 activity in crude extracts. In the E1 assay an artificial eiectronacceptor has to be added. We use dichlorophenolindophenol (DCPIP) instead of the often applied ferricyanide. DCPIP shows no inhibitory effects on E1 activity. E1 activity appears to be dependent on the cofactors thiaminepyrophosphate and magnesium and is completely inhibited by fluoropyruvate. The method can be applied to frozen or fresh small muscle samples, obtained by needle or open biopsy.
We established a difference in E1 activity between 600 g supernatant and horrogenate of skeletal muscle tissue. The measurement of E1 activity in supernatant from frozen muscle appeared to be unreliable. Control values in homogenate, 515-2080 nmol.hr−1.g−1. muscle, are higher or the same as those reported by others. With the new assay we identified two patients with a E1 deficiency.
The method appears also suitable for measurement of E1 activity in chorionic villi.
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Van Laack, R., Ruitenbeek, W., Trijbels, F. et al. 149 ESTIMATION OF PYRUVATE DEHYDROGENASE (E1) ACTIVITY IN HUMAN SKELETAL MUSCLE. Pediatr Res 20, 1059 (1986). https://doi.org/10.1203/00006450-198610000-00204
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DOI: https://doi.org/10.1203/00006450-198610000-00204
