1989 Fiscal Year Final Research Report Summary
Mechanisms of the intracellular targeting of lysosomal cysteine proteinase
| Project/Area Number |
63480128
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
General medical chemistry
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| Research Institution | The University of Tokushima |
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Principal Investigator |
KATUNUMA N Institute for Enzyme Research, The University of Tokushima, Professor, 酵素科学研究センター, 教授 (50035375)
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| Co-Investigator(Kenkyū-buntansha) |
II K The University of Tokushima, School of Medicine Assistant Professor, 医学部, 助教授 (50035507)
KOMINAMI E Juntendo University School of Medicine, Professor, 医学部, 教授 (10035496)
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| Project Period (FY) |
1988 – 1989
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| Keywords | Cathepsin / Lysosomes / Intracellular processing / Macrophages / cDNA cloning |
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| Research Abstract |
Inatracellular targeting of lysosomal cysteine proteinases to lysosomes was studied with cultured macrophages. Cathepsins B, H and L were found to be as proenzymes in Golgi apparatus, and then as the single chain mature enzymes in lysosomes. Metallo-proteinase inhibitors and pepstatin retarded the conversion of proenzymes to mature enzymes. Short pulse-chase experiment showed the detection of intermediates during the conversion, suggesting that different proteinases operate in the conversion process. cDNA cloning and sequencing of cathepsin H and L were carried out and amino acid sequences of pre-propeptides of enzymes were deduced. Immunoelectron microscopic studies with antibodies specific for procathepsins showed that mature enzymes were detected in lysosomes, and that proenzymes were localized in Golgi apparatus. It is under investigation what forms of enzymes are present in prelysosomes. More than 30% of procathepsins synthesized was excreted to the medium without reaching to lysosomes. Morphologically, small vesides labeled by anticathepsin L were detected, suggesting that those are in secretory route.
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Research Products
(14 results)
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[Publications] M. Watanabe, T. Watanabe, Y. Ishii, M. Matsuba, S. Kimura, E. Kominami, N. Katunuma and Y. Uchiyama: "Immunohistochemical localization of cathepsin B, H and their endogenous inhibitor, cystatin beta in islet endocrine cells of the rat pancreas" J. Histochem. Cytochem. 36-7, 783-791, 1988.
Description
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