{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,10,14]],"date-time":"2023-10-14T08:14:53Z","timestamp":1697271293623},"reference-count":29,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2006,10,2]],"date-time":"2006-10-02T00:00:00Z","timestamp":1159747200000},"content-version":"vor","delay-in-days":1,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Clin Experimental Allergy"],"published-print":{"date-parts":[[2006,10]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p><jats:bold>Background\n\t\t\t\t\t<\/jats:bold> Mast cell mediators are believed to play a central role in inflammatory lung disorders such as human allergic and occupational asthma. Equine heaves is characterized by reversible neutrophilic airway inflammation and airway obstruction, primarily due to bronchospasm and mucus hypersecretion, following exposure of susceptible horses to organic stable dusts. As such, heaves shares many similarities with human occupational dust\u2010induced asthma and therefore it is proposed that mast cells may also be implicated in the pathogenesis of heaves. Tryptase, a mast cell\u2010specific proteinase, can be used as an indicator of biological mast cell activity.<\/jats:p><jats:p><jats:bold>Objective\n\t\t\t\t\t<\/jats:bold> The aim of this study was to determine the cDNA sequence of equine tryptase and to investigate its substrate specificity in order to rationalize its enzymatic activity.<\/jats:p><jats:p><jats:bold>Methods\n\t\t\t\t\t<\/jats:bold> RT\u2010PCR cloning was used to sequence equine tryptase. Substrate specificity of equine tryptase was investigated using arginine and lysine containing substrates.<\/jats:p><jats:p><jats:bold>Results\n\t\t\t\t\t<\/jats:bold> The cDNA and deduced amino acid (Aa) sequences for equine tryptase shared strong identity with other tryptases. Unusually for a trypsin\u2010like proteinase however, equine tryptase has alanine at residue 216, rather than glycine, which confers increased arginine substrate specificity <jats:italic>in vitro<\/jats:italic> and may restrict fibrinogenolysis <jats:italic>in vivo<\/jats:italic>.<\/jats:p><jats:p><jats:bold>Conclusion\n\t\t\t\t\t<\/jats:bold> Cloning and sequencing of the mast cell proteinase equine tryptase will allow molecular probing of its expression in the lung of control and heaves\u2010affected horses. Further work is warranted to determine the biological relevance of the unique alanine 216 substitution in the molecular sequence of the equine tryptase substrate\u2010binding pocket.<\/jats:p>","DOI":"10.1111\/j.1365-2222.2006.02571.x","type":"journal-article","created":{"date-parts":[[2006,10,2]],"date-time":"2006-10-02T14:33:17Z","timestamp":1159799597000},"page":"1303-1309","source":"Crossref","is-referenced-by-count":2,"title":["cDNA cloning and substrate specificity of equine tryptase, a possible mediator in equine heaves"],"prefix":"10.1111","volume":"36","author":[{"given":"K. J.","family":"Dacre","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S. M.","family":"McAleese","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"P.","family":"Knight","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"B. 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Crystallographic studies of substrates and inhibitors bound to the active site of [alpha]-chymotrypsin"},{"key":"e_1_2_7_21_2","first-page":"2291","article-title":"Human mast cell tryptase fibrinogenolysis","volume":"37","author":"Thomas VA","year":"1998","journal-title":"kinetics, anticoagulation mechanism, and cell adhesion disruption"},{"key":"e_1_2_7_22_2","doi-asserted-by":"crossref","first-page":"719","DOI":"10.1042\/bj3230719","article-title":"Sheep mast cell proteinase\u20101, a serine proteinase with both tryptase\u2010 and chymase\u2010like properties, is inhibited by plasma proteinase inhibitors and is mitogenic for bovine pulmonary artery fibroblasts","volume":"323","author":"Pemberton AP","year":"1997","journal-title":"Biochem J"},{"key":"e_1_2_7_23_2","doi-asserted-by":"crossref","first-page":"3875","DOI":"10.1016\/S0021-9258(18)34864-6","article-title":"Computer\u2010generated models of blood coagulation factor Xa, factor IXa, and thrombin based upon structural homology with other serine proteases","volume":"257","author":"Furie B","year":"1982","journal-title":"J Biol Chem"},{"key":"e_1_2_7_24_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00005a004"},{"key":"e_1_2_7_25_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI114284"},{"key":"e_1_2_7_26_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.50.31885"},{"key":"e_1_2_7_27_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcb.240330307"},{"key":"e_1_2_7_28_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi015662v"},{"key":"e_1_2_7_29_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.274.28.19670"},{"key":"e_1_2_7_30_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI118337"}],"container-title":["Clinical &amp; Experimental Allergy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fj.1365-2222.2006.02571.x","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/j.1365-2222.2006.02571.x","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,13]],"date-time":"2023-10-13T19:10:55Z","timestamp":1697224255000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/j.1365-2222.2006.02571.x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2006,10]]},"references-count":29,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2006,10]]}},"alternative-id":["10.1111\/j.1365-2222.2006.02571.x"],"URL":"https:\/\/doi.org\/10.1111\/j.1365-2222.2006.02571.x","archive":["Portico"],"relation":{},"ISSN":["0954-7894","1365-2222"],"issn-type":[{"value":"0954-7894","type":"print"},{"value":"1365-2222","type":"electronic"}],"subject":[],"published":{"date-parts":[[2006,10]]}}}