Immuno-electron microscopy on the morphological marker enzymes and proteins for subcompartments of cell organellae.

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Morphological marker enzymes and proteins of subcompartments in cell organellae have been investigated, from two view points of the ultrastructure and protein molecules by immuno-electron microscopy.1.New findings were obtained mainly on the paroxysm, the mitochoindria and the nucleus.(1) Urate oxidase, which has been considered to be a common constituent of nucleoid core of mammalian hepatic peroxisomes, was demonstrated to be a morphological marker enzyme of vertebrate hepatic peroxisomes.(2) The process and functional aspect of the finding (1) was further investigated using cultured cells as materials.(3) Fatty acid beta-oxidation enzymes have been show to be commonly localized in the mitochondrial cristae, and suggested to be its marker.(4) Allantoinase was show to be a mitochondrial enzyme and as demonstrated to be a morphological marker enzyme of mitochondrial matrix.(5) A calcineurin isoform, CAP 50 protein and PCNA/cyclin were shown to be marker proteins for the nucleus.2.Problems in the research. Unexpectedly long time was needed to purify enzymes and proteins of endoplasmic reticulum, Golgi apparatus, and lysosomes. Even when they were purified, unexpected non-specific reaction existed in the antibodies. These two problem resulted in unsatisfactory results in these three cell organellas.3.Problem solutions. In order to achieve this object, immunoelectron microscopic studies employing the antibodies for synthetic polypeptides is being started.