| Binding of granzyme B in the nucleus of target cells. Recognition of an 80-kilodalton protein. | |
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MedLine Citation:
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PMID: 8626587 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Granzyme B (cytotoxic cell proteinase 1) is a serine proteinase that has been implicated in cytotoxic T lymphocyte-induced apoptosis. In order to understand how granzyme B is involved in mechanisms of target cell destruction, characterization and identification of substrates are required. We have developed an in situ binding assay using permeabilized cells and recombinant granzyme B that allows us to visualize potential substrates after immunostaining with anti-granzyme B antiserum. Confocal laser scanning microscopy and immunoelectron microscopic analyses demonstrate that granzyme B recognizes a nuclear substrate. The labeling pattern observed corresponds with regions of positive staining with uranyl acetate which binds to heterochromatin in the nucleus. Positive labeling of target cells with granzyme B is dependent on the presence of a catalytically active proteinase, since an inactive proenzyme form of granzyme B fails to give rise to any binding in the target cells. Far-Western blotting and immunoprecipitation of subcellular fractions of target cells have shown that the putative substrate of catalytically active granzyme B is an 80-kDa nuclear protein. Minor cytosolic bands of 50 and 94 kDa are also observed. A cytoplasmic band of 69 kDa is detected by both active and zymogen forms of granzyme B. |
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Authors:
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M J Pinkoski; U Winkler; D Hudig; R C Bleackley |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: The Journal of biological chemistry Volume: 271 ISSN: 0021-9258 ISO Abbreviation: J. Biol. Chem. Publication Date: 1996 Apr |
Date Detail:
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Created Date: 1996-06-21 Completed Date: 1996-06-21 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 2985121R Medline TA: J Biol Chem Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 10225-9 Citation Subset: IM |
Affiliation:
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Department of Biochemistry, University of Alberta, Edmonton, Alberta, T6G 2H7, Canada. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Compartmentation Cell Nucleus / metabolism* Enzyme Precursors / metabolism Fluorescent Antibody Technique, Indirect Granzymes Leukemia L1210 Microscopy, Electron, Scanning Molecular Weight Nuclear Proteins / chemistry, metabolism* Protein Binding Recombinant Proteins Serine Endopeptidases / metabolism* Substrate Specificity T-Lymphocytes, Cytotoxic / enzymology* |
| Chemical | |
Reg. No./Substance:
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0/Enzyme Precursors; 0/Nuclear Proteins; 0/Recombinant Proteins; EC 3.4.21.-/Granzymes; EC 3.4.21.-/Serine Endopeptidases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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