{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T04:37:17Z","timestamp":1768451837341,"version":"3.49.0"},"reference-count":8,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2002,3,13]],"date-time":"2002-03-13T00:00:00Z","timestamp":1015977600000},"content-version":"vor","delay-in-days":12,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J of Applied Toxicology"],"published-print":{"date-parts":[[2002,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>One of the mechanisms of the skin blistering effect (vesication) of sulfur mustard (bis\u2010(2\u2010chloroethyl)sulfide, HD) is believed to be via the stimulation of specific protease(s) at the dermal\u2013epidermal junction. Cultured normal human epidermal keratinocytes (NHEK) were used as a model to study and characterize protease stimulated by the mustards 2\u2010chloroethyl ethyl sulfide (CEES), 2\u2010chloro\u2010<jats:italic>N<\/jats:italic>\u2010(2\u2010chloroethyl)\u2010<jats:italic>N<\/jats:italic>\u2010methylethanamine hydrochloride (nitrogen mustard, HN<jats:sub>2<\/jats:sub>) and HD. The results obtained using a chromozym (TRY) peptide substrate protease assay revealed the optimum mustard concentrations and time for protease stimulation to be about 200 \u00b5M (CEES), 100 \u00b5M(HN<jats:sub>2<\/jats:sub>) and 100 \u00b5M (HD) and 16 h. The mustard\u2010stimulated protease was membrane bound and was inhibited by adding a Ca<jats:sup>2+<\/jats:sup> chelator (either 2 mM EGTA (ethylene glycol\u2010bis(amino ethyl ether) N,N,N\u2032,N\u2032 tetraacetic acid) or 50 \u00b5M BAPTA AM (1,2\u2010bis(z\u2010aminophenoxy)ethane\u2010N,N,N\u2032,N\u2032\u2010tetraacetic acid, tetraacetoxy methyl ester) alone or in combination), a serine protease inhibitor diisopropyl fluoro\u2010phosphate (DFP, 1 mM), or a protein synthesis inhibitor cycloheximide (35 \u00b5M) in the extracellular medium. These results suggest that mustard toxicity may involve the stimulation of trypsin\/chymotrypsin\u2010like serine protease, dependent on Ca<jats:sup>2+<\/jats:sup> and new protein synthesis. Protein purification by gel exclusion and hydrophobic chromatography produced a 70\u201380 kDa protease, which had an amino acid sequence homologous with a mammalian\u2010type bacterial serine endopeptidase. Based on this information, research is in progress to identify the protease stimulated by HD in NHEK and to determine whether its inhibitors are useful as prospective antivesicant drugs. Copyright \u00a9 2002 John Wiley &amp; Sons, Ltd.<\/jats:p>","DOI":"10.1002\/jat.829","type":"journal-article","created":{"date-parts":[[2002,8,25]],"date-time":"2002-08-25T19:42:44Z","timestamp":1030304564000},"page":"139-140","source":"Crossref","is-referenced-by-count":16,"title":["Sulfur mustard\u2010stimulated protease: a target for antivesicant drugs"],"prefix":"10.1002","volume":"22","author":[{"given":"P.","family":"Ray","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"A. K.","family":"Chakrabarti","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C. A.","family":"Broomfield","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R.","family":"Ray","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2002,3,13]]},"reference":[{"key":"e_1_2_1_2_2","first-page":"A 828","article-title":"Increased proteolytic activity in human epithelial cells following exposure to sulfur mustard","volume":"5","author":"Smith WJ","year":"1991","journal-title":"FASEB J."},{"key":"e_1_2_1_3_2","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1007\/BF00250978","article-title":"Effect of sulfur mustard exposure on protease activity in human peripheral blood lymphocytes","volume":"7","author":"Cowan FM","year":"1991","journal-title":"Cell Biol. Toxicol."},{"key":"e_1_2_1_4_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0029(19970501)37:3<221::AID-JEMT7>3.0.CO;2-Q"},{"key":"e_1_2_1_5_2","first-page":"433","volume-title":"Proc. 1989 Medical Defense Bioscience Review","author":"Broomfield CA","year":"1989"},{"key":"e_1_2_1_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/227680a0"},{"key":"e_1_2_1_7_2","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/0003-2697(76)90527-3","article-title":"A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein\u2013dye binding","volume":"72","author":"Bradford M","year":"1976","journal-title":"Ann. Biochem."},{"key":"e_1_2_1_8_2","unstructured":"RayP. Protein biomarker for mustard chemical injury. US Patent 08\/855 13 May1997; 268."},{"key":"e_1_2_1_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-2952(98)00160-9"}],"container-title":["Journal of Applied Toxicology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fjat.829","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jat.829","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T10:18:23Z","timestamp":1760523503000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/jat.829"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2002,3]]},"references-count":8,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2002,3]]}},"alternative-id":["10.1002\/jat.829"],"URL":"https:\/\/doi.org\/10.1002\/jat.829","archive":["Portico"],"relation":{},"ISSN":["0260-437X","1099-1263"],"issn-type":[{"value":"0260-437X","type":"print"},{"value":"1099-1263","type":"electronic"}],"subject":[],"published":{"date-parts":[[2002,3]]}}}