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PMID:9681477

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Citation

Vogel, R, Jennemann, G, Seitz, J, Wiesinger, H and Hamprecht, B (1998) Mitochondrial malic enzyme: purification from bovine brain, generation of an antiserum, and immunocytochemical localization in neurons of rat brain. J. Neurochem. 71:844-52

Abstract

To elucidate the cellular location of mitochondrial malic enzyme in brain, immunocytochemical studies were performed. For this purpose, mitochondrial malic enzyme was purified to apparent homogeneity from bovine brain and used for the immunization of rabbits. Subjecting the antiserum to affinity purification on immobilized antigen as an absorbent yielded a purified immunoreactive antibody preparation, which was characterized by probing cytosolic and mitochondrial fractions of bovine and rat brain in western blotting. As neither cross-reactivity with cytosolic malic enzyme nor immunoreactivity against other proteins could be observed, the antibody preparation was found suitable for immunocytochemistry. By using sections of perfusion-fixed rat brain, considerable resolution was achieved at the light-microscopic level. Distinct and specific staining of neurons was observed; in contrast, no staining of astrocytes and possibly unspecific staining within the nuclei of oligodendrocytes were obtained. From these data, it is concluded that mitochondrial malic enzyme is located in neurons; however, in astrocytes, the enzyme appears to be either lacking or present at a much lower level. A protective role against oxidative stress in neurons is proposed for mitochondrial malic enzyme.

Links

PubMed

Keywords

Animals; Antibodies/isolation & purification; Blotting, Western; Cattle; Enzyme-Linked Immunosorbent Assay; Glutathione/metabolism; Hippocampus/cytology; Hippocampus/enzymology; Malate Dehydrogenase/analysis; Malate Dehydrogenase/immunology; Malate Dehydrogenase/metabolism; Mitochondria/enzymology; NADP/metabolism; Neocortex/cytology; Neocortex/enzymology; Neurons/enzymology; Oxidative Stress/physiology; Rabbits; Rats

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

RAT:MAOX

GO:0005739: mitochondrion

ECO:0000314:

C

See Figure 2.

complete
CACAO 5782

RAT:MAOX

part_of

GO:0005739: mitochondrion

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

BOVIN:F1N3V0

GO:0005739: mitochondrion

ECO:0000314:

C

See figure 2 lane C for bovine brain mitochondrial fraction

complete
CACAO 5789

BOVIN:F1N3V0

part_of

GO:0005739: mitochondrion

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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