The subcellular distribution of
2-oxoglutarate:
glyoxylate carboligase was investigated in a normal human liver, a liver from a patient with
pyridoxine-resistant
primary hyperoxaluria type I and rat livers subjected to various degrees and types of
trauma. On continuous
sucrose gradients most of the carboligase fractionated with a peak equilibrium density of 1.19-1.20 g/cm3 and paralleled the distribution of the major peaks of
monoamine oxidase,
glutamate dehydrogenase and
cytochrome oxidase and can be considered to be mitochondrial. Various proportions of the carboligase and mitochondrial marker
enzymes were found to be 'extramitochondrial' (at or near the top of the
sucrose gradients), depending on the liver source and the severity of
trauma to which they were subjected. Carboligase,
monoamine oxidase (outer membrane marker) and
glutamate dehydrogenase (matrix marker) were released from mitochondria by the homogenization and centrifugation procedures, to the extent of 19.9%, 32.4% and 11.5% respectively in hyperoxaluric liver, 12.5%, 17.9% and 8.2% in normal human liver and 3.0%, 4.9% and 3.8% in control rat liver. The proportion of extramitochondrial
cytochrome oxidase (inner membrane marker) was virtually undetectable in both human and rat livers. However, sonication of rat liver homogenates or the addition of the
detergent Triton X-100 caused a massive release of all four
enzymes. The extramitochondrial carboligase was probably in the form of a free
protein of very high molecular weight or aggregate, rather than associated with a mitochondrion-derived organelle. Subfractionation of a rat liver mitochondrial preparation indicated that most of the carboligase activity paralleled activities of
2-oxoglutarate decarboxylase,
citrate synthase and
glutamate dehydrogenase and was probably located in the matrix.(ABSTRACT TRUNCATED AT 250 WORDS)