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Determination of Compartmented Metabolite Pools by a Combination of Rapid Fractionation of Oat Mesophyll Protoplasts and Enzymic Cycling ¹

Institut für Biologie I, Universität Tübingen, Auf der Morgenstelle 1, D-7400 Tübingen 1, Institut für Botanik, Technische Universität München, Arcisstrasse 21, D-8000 München 2, Federal Republic of Germany

In vivo pool sizes of a range of metabolites have been determined in subcellular fractions of darkened and illuminated mesophyll protoplasts of Avena sativa L. These estimations were made by combining a method of rapid protoplast fractionation with enzymic cycling techniques. Results are given for reduced and oxidized pyridine nucleotides, triose phosphates, 3-phosphoglycerate, inorganic phosphate, aspartate, malate, oxaloacetate, glutamate, 2-oxoglutarate, and citrate, from chloroplasts, mitochondria, and a fraction representing the remainder of the protoplast. The results indicate distinct differences of compartmented levels of certain metabolites between darkened and illuminated protoplasts.

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