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PMID: 6289811 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Limitations of commonly used spectrophotometric assay methods for phosphoenolypyruvate carboxykinase activity in crude extracts of muscle.

The Biochemical journal ·Vol. 206 ·No. 1 ·1982-07-15 ·Pages 147-52

Duff DA, Snell K

Abstract

Phosphoenolpyruvate carboxykinase activity in crude extracts of muscle has frequently been determined by using a continuous spectrophotometric method, which is shown to grossly overestimate enzyme activity. NADH oxidation attributed to phosphoenolpyruvate carboxykinase activity in the assay is due to lactate production. Under the normal assay conditions. Na+ ions stimulate pyruvate kinase, providing pyruvate for lactate formation by lactate dehydrogenase and sufficiently to account for most of the observed NADH oxidation.

MeSH Terms
Animals Bicarbonates/pharmacology Carboxy-Lyases/metabolism In Vitro Techniques L-Lactate Dehydrogenase/metabolism Lactates/metabolism Lactic Acid Male Muscles/enzymology NAD/metabolism Phosphoenolpyruvate Carboxylase/metabolism Pyruvate Kinase/metabolism Rats Rats, Inbred Strains Sodium Bicarbonate Sodium Chloride/pharmacology Spectrophotometry/methods
Chemicals
Bicarbonates Lactates NAD Lactic Acid Sodium Chloride Sodium Bicarbonate L-Lactate Dehydrogenase Pyruvate Kinase Carboxy-Lyases Phosphoenolpyruvate Carboxylase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Duff D A
Snell K
References (19)
19 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2060147
Published
1982-07-15
Pages
147-52
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1158560
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2060147 PMC Full Text View journal details
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