{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T17:01:12Z","timestamp":1760979672920},"reference-count":0,"publisher":"Rockefeller University Press","issue":"6","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[1994,12,1]]},"abstract":"<jats:p>We report the purification of a protease from tumor cells undergoing apoptosis that is involved in activating DNA fragmentation. Initial studies revealed that two inhibitors of serine proteases, N-1-tosylamide-2-phenylethylchloromethyl ketone and carbobenzoxy-Ala-Ala-borophe (DK120), suppressed tumor necrosis factor or ultraviolet (UV) light-induced DNA fragmentation in the U937 histiocytic lymphoma as well as UV light-induced DNA fragmentation in the BT-20 breast carcinoma, HL-60 myelocytic leukemia, and 3T3 fibroblasts. The protease was purified by affinity chromatography with DK120 as ligand and showed high activity on a synthetic substrate preferred by elastase-like enzymes (Ala-Ala-Pro-Val p-nitroanilide), but was inactive on the trypsin substrate, N-alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester, or the chymotrypsin substrate, Ala-Ala-Pro-Phe p-nitroanilide. The activity of the DK120-binding protease purified from U937 cells undergoing apoptosis was increased approximately 10-fold over that recovered from normal cells. Further purification to homogeneity by heparin-Sepharose affinity chromatography followed by reverse phase high-performance liquid chromatography revealed a single band of 24 kD on a silver-stained sodium dodecyl sulfate gel. In addition to protease activity, the purified enzyme induced DNA fragmentation into multiples of 180 basepairs in isolated U937 nuclei. These findings suggest the 24-kD protease is a novel enzyme that activates DNA fragmentation in U937 cells undergoing apoptosis.<\/jats:p>","DOI":"10.1084\/jem.180.6.2113","type":"journal-article","created":{"date-parts":[[2004,6,24]],"date-time":"2004-06-24T07:56:10Z","timestamp":1088063770000},"page":"2113-2123","update-policy":"http:\/\/dx.doi.org\/10.1084\/jem.crossmarkpolicy","source":"Crossref","is-referenced-by-count":57,"title":["Purification of a 24-kD protease from apoptotic tumor cells that activates DNA fragmentation."],"prefix":"10.1084","volume":"180","author":[{"given":"S C","family":"Wright","sequence":"first","affiliation":[{"name":"Palo Alto Institute of Molecular Medicine, Mountain View, California 94043."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Q S","family":"Wei","sequence":"additional","affiliation":[{"name":"Palo Alto Institute of Molecular Medicine, Mountain View, California 94043."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J","family":"Zhong","sequence":"additional","affiliation":[{"name":"Palo Alto Institute of Molecular Medicine, Mountain View, California 94043."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"H","family":"Zheng","sequence":"additional","affiliation":[{"name":"Palo Alto Institute of Molecular Medicine, Mountain View, California 94043."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"D H","family":"Kinder","sequence":"additional","affiliation":[{"name":"Palo Alto Institute of Molecular Medicine, Mountain View, California 94043."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J W","family":"Larrick","sequence":"additional","affiliation":[{"name":"Palo Alto Institute of Molecular Medicine, Mountain View, California 94043."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"291","published-online":{"date-parts":[[1994,12,1]]},"container-title":["The Journal of experimental medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jem\/article-pdf\/180\/6\/2113\/1676504\/2113.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jem\/article-pdf\/180\/6\/2113\/1676504\/2113.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,25]],"date-time":"2023-07-25T06:32:53Z","timestamp":1690266773000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jem\/article\/180\/6\/2113\/25678\/Purification-of-a-24-kD-protease-from-apoptotic"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1994,12,1]]},"references-count":0,"journal-issue":{"issue":"6","published-print":{"date-parts":[[1994,12,1]]}},"URL":"https:\/\/doi.org\/10.1084\/jem.180.6.2113","relation":{},"ISSN":["0022-1007","1540-9538"],"issn-type":[{"value":"0022-1007","type":"print"},{"value":"1540-9538","type":"electronic"}],"subject":[],"published":{"date-parts":[[1994,12,1]]}}}