Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
9
|
| pubmed:dateCreated |
1994-10-18
|
| pubmed:abstractText |
We have partially characterized the granules of the human NK cell line, YT-INDY, and assessed granule-mediated lysis and DNA fragmentation of assorted targets. Biochemical studies demonstrated significant quantities of granzyme B (asp-ase) and a heretofore undescribed chymase but no tryptase (i.e., granzyme A or 3) or distinct met-ase. YT-INDY expressed mRNA for granzyme B, perforin and CCPX. The existence of perforin was confirmed by immunoblot. The granules lysed both human and murine NK-sensitive and NK-resistant targets. YT-INDY and NK3.3, two human cytotoxic cells, were also lysed. EGTA reduced lysis by only 50%, suggesting that a perforin-independent lytic pathway is associated with the granules. In addition, 4-(2-aminoethyl) benzenesulfonylfluoride hydrochloride (AEBSF), an inhibitor that selectively blocked the chymase and 3,4-dichloroisocoumarin (DCI), an inhibitor that inactivated both chymase and asp-ase activities, marginally affected lysis. By gel electrophoresis and 125I-labeled deoxyuridine release assay, only murine cells (SP2/0 and YAC-1) underwent DNA fragmentation, and cleavage was completely inhibited by DCI, whereas EGTA, AEBSF and aurintricarboxylic acid (ATA) had no effect. The results, therefore, underscore the central role of granzyme B in granule-mediated DNA fragmentation, emphasize that the protease acts via an ATA-resistant endonuclease pathway and stress that nucleolysis does not invariably accompany granule-mediated cytolysis. Finally, ATA inhibited the asp-ase activity of isolated but not granule-associated granzyme B. ATA, therefore, is not a specific endonuclease inhibitor and results obtained with ATA should be viewed cautiously.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/3,4-dichloroisocoumarin,
http://linkedlifedata.com/resource/pubmed/chemical/4-(2-aminoethyl)benzenesulfonylfluor...,
http://linkedlifedata.com/resource/pubmed/chemical/Aurintricarboxylic Acid,
http://linkedlifedata.com/resource/pubmed/chemical/Chymases,
http://linkedlifedata.com/resource/pubmed/chemical/Coumarins,
http://linkedlifedata.com/resource/pubmed/chemical/DNA,
http://linkedlifedata.com/resource/pubmed/chemical/Egtazic Acid,
http://linkedlifedata.com/resource/pubmed/chemical/GZMB protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Granzymes,
http://linkedlifedata.com/resource/pubmed/chemical/Gzmb protein, mouse,
http://linkedlifedata.com/resource/pubmed/chemical/Serine Endopeptidases,
http://linkedlifedata.com/resource/pubmed/chemical/Sulfones
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Sep
|
| pubmed:issn |
0014-2980
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
24
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
2073-80
|
| pubmed:dateRevised |
2006-11-15
|
| pubmed:meshHeading |
pubmed-meshheading:8088328-Animals,
pubmed-meshheading:8088328-Aurintricarboxylic Acid,
pubmed-meshheading:8088328-Blotting, Northern,
pubmed-meshheading:8088328-Blotting, Western,
pubmed-meshheading:8088328-Cell Survival,
pubmed-meshheading:8088328-Chymases,
pubmed-meshheading:8088328-Coumarins,
pubmed-meshheading:8088328-Cytoplasmic Granules,
pubmed-meshheading:8088328-Cytotoxicity, Immunologic,
pubmed-meshheading:8088328-DNA,
pubmed-meshheading:8088328-Egtazic Acid,
pubmed-meshheading:8088328-Granzymes,
pubmed-meshheading:8088328-Humans,
pubmed-meshheading:8088328-Killer Cells, Natural,
pubmed-meshheading:8088328-Mice,
pubmed-meshheading:8088328-Serine Endopeptidases,
pubmed-meshheading:8088328-Sulfones,
pubmed-meshheading:8088328-Tumor Cells, Cultured
|
| pubmed:year |
1994
|
| pubmed:articleTitle |
Human granzyme B is essential for DNA fragmentation of susceptible target cells.
|
| pubmed:affiliation |
Department of Medicine, Indiana University School of Medicine, Indianapolis.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|