An immobilized three enzyme system

A model for microenvironmental compartmentation in mitochondria

P. Srere, B. Mattiasson, K. Mosbach

Research output: Contribution to journalArticle

151 Citations (Scopus)

Abstract

An immobilized three enzyme system, malate dehydrogenase (EC 1.1.1.37) - citrate synthase (EC 4.1.3.7) - lactate dehydrogenase (EC 1.1.1.27), was investigated as a model for the rate of oxalacetate production and utilization in mitochondria. Lactate dehydrogenase is included to mimic the NADH utilizing system of mitochondria. This three enzyme system was immobilized in three different ways on Sephadex G 50 (surface coupling), on Sepharose 4B (internal external coupling), and entrapped in polyacrylamide gel. The rate of citrate production from malate, NAD+, and acetyl CoA was determined continuously in a flow system. Up to about 100% rate enhancements were observed when the immobilized system was compared to identical systems of free enzyme. An even more pronounced increase of rate of up to about 400% compared to the soluble system was measured after addition of pyruvate (to reoxidize formed NADH). These results are interpreted in relation to microenvironmental changes of oxalacetate production and the possible organization of enzymes of the Krebs cycle.

Original languageEnglish
Pages (from-to)2534-2538
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume70
Issue number9
Publication statusPublished - 1973

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Immobilized Enzymes
NAD
Mitochondria
L-Lactate Dehydrogenase
Enzymes
Citrate (si)-Synthase
Malate Dehydrogenase
Acetyl Coenzyme A
Citric Acid Cycle
Pyruvic Acid
Citric Acid
Sepharose

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

An immobilized three enzyme system : A model for microenvironmental compartmentation in mitochondria. / Srere, P.; Mattiasson, B.; Mosbach, K.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 70, No. 9, 1973, p. 2534-2538.

Research output: Contribution to journalArticle

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