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Research ArticleMetabolism and Enzymology
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Characterization of Biotin and 3-Methylcrotonyl-Coenzyme A Carboxylase in Higher Plant Mitochondria

Pierre Baldet, Claude Alban, Stella Axiotis, Roland Douce
Pierre Baldet
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Claude Alban
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Stella Axiotis
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Roland Douce
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Published June 1992. DOI: https://doi.org/10.1104/pp.99.2.450

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  • © 1992 American Society of Plant Biologists

Abstract

Mitochondria from green pea (Pisum sativum) leaves were purified free of peroxisomes and chlorophyll contamination and examined for their biotin content. The bulk of the bound biotin detected in plant mitochondria was shown to be associated with the matrix space to a concentration of about 13 micromolar, and no free biotin was detected. Western blot analysis of mitochondrial polypeptides using horseradish peroxidase-labeled streptavidin revealed a unique biotin-containing polypeptide with a molecular weight of 76,000. This polypeptide was implicated as being the biotinylated subunit of 3-methylcrotonyl-coenzyme A (CoA) carboxylase. Fractionation of pea leaf protoplasts demonstrated that this enzyme activity was located largely in mitochondria. The 3-methylcrotonyl-CoA carboxylase activity was latent when assayed in isotonic media. The majority of the enzyme activity was found in the soluble matrix of mitochondria. Maximal 3-methylcrotonyl-CoA carboxylase activity was found at pH 8.3 in the presence of Mg2+. Kinetic constants (apparent Km values) for the enzyme substrates were: 3-methylcrotonyl-CoA, 0.05 millimolar; ATP, 0.16 millimolar; HCO3−, 2.2 millimolar. The involvement of 3-methylcrotonyl-CoA carboxylase in the leucine degradation pathway in plant mitochondria is proposed.

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Characterization of Biotin and 3-Methylcrotonyl-Coenzyme A Carboxylase in Higher Plant Mitochondria
Pierre Baldet, Claude Alban, Stella Axiotis, Roland Douce
Plant Physiology Jun 1992, 99 (2) 450-455; DOI: 10.1104/pp.99.2.450

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Characterization of Biotin and 3-Methylcrotonyl-Coenzyme A Carboxylase in Higher Plant Mitochondria
Pierre Baldet, Claude Alban, Stella Axiotis, Roland Douce
Plant Physiology Jun 1992, 99 (2) 450-455; DOI: 10.1104/pp.99.2.450
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Plant Physiology
Vol. 99, Issue 2
June 1992
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More in this TOC Section

  • Tomato Fruit Polygalacturonase Isozyme 1 (Characterization of the [beta] Subunit and Its State of Assembly in Vivo)
  • A Novel Metabolic Pathway for Indole-3-Acetic Acid in Apical Shoots of Populus tremula (L.) x Populus tremuloides (Michx.)
  • Biosynthesis of Cardiolipin in Plant Mitochondria
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