{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T08:58:06Z","timestamp":1789376286188,"version":"build-2803163510"},"reference-count":19,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2008,12,31]],"date-time":"2008-12-31T00:00:00Z","timestamp":1230681600000},"content-version":"vor","delay-in-days":5570,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[1993,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Echistatin is the smallest member of the disintegrin family of snake venom proteins, containing four disulfides in a peptide chain of 49 residues. Partial assignment of disulfides has been made previously by NMR and chemical approaches. A full assignment was made by a newly developed chemical approach, using partial reduction with tris\u2010(2\u2010carboxyethyl)\u2010phosphine at acid pH. Reduction proceeded in a stepwise manner at pH 3, and the intermediates were isolated by high performance liquid chromatography. Alkylation of free thiols, followed by sequencer analysis, enabled all four bridges to be identified: (1) at 20 \u00b0C a single bridge linking Cys 2\u2010Cys 11 was broken, giving a relatively stable intermediate; (2) with further treatment at 41 \u00b0C the bridges Cys 7\u2010Cys 32 and Cys 8\u2010Cys 37 became accessible to the reagent and were reduced at approx. equal rates; (3) the two bicyclic peptides produced in this manner were less stable and could be reduced at 20 \u00b0C to a peptide that retains a single bridge linking Cys 20\u2010Cys 39; and (4) the monocyclic peptide can be reduced to the linear molecule at 20 \u00b0C. Some disulfide exchange occurred during alkylation of the bicyclic intermediates, but results unambiguously show the pattern to be [2\u201311; 7\u201332; 8\u201337; 20\u201339]. A comparison is made with kistrin, a longer disintegrin whose disulfide structure has been proposed from NMR analysis.<\/jats:p>","DOI":"10.1002\/pro.5560021018","type":"journal-article","created":{"date-parts":[[2009,2,10]],"date-time":"2009-02-10T00:01:22Z","timestamp":1234224082000},"page":"1749-1755","source":"Crossref","is-referenced-by-count":41,"title":["Echistatin disulfide bridges: Selective reduction and linkage assignment"],"prefix":"10.1002","volume":"2","author":[{"given":"William R.","family":"Gray","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2008,12,31]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00052a036"},{"key":"e_1_2_1_3_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1862345"},{"key":"e_1_2_1_4_1","doi-asserted-by":"publisher","DOI":"10.1021\/jf00014a037"},{"key":"e_1_2_1_5_1","doi-asserted-by":"publisher","DOI":"10.1021\/jo00008a014"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00235a016"},{"key":"e_1_2_1_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(92)80797-K"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00114a004"},{"key":"e_1_2_1_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1991.tb16379.x"},{"key":"e_1_2_1_10_1","doi-asserted-by":"publisher","DOI":"10.1093\/protein\/5.6.473"},{"key":"e_1_2_1_11_1","unstructured":"Craig A.G. Mahe E. Fischer W.H. Gray W.R. andRivier J.E.(1993).Strategy to determine disulfide bridge arrangements by selective reduction of cystines using TCEP. Abstract 41st Annual Conference on Mass Spectrometry San Francisco."},{"key":"e_1_2_1_12_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1991.tb16378.x"},{"key":"e_1_2_1_13_1","doi-asserted-by":"crossref","first-page":"19827","DOI":"10.1016\/S0021-9258(19)77710-2","article-title":"Echistatin: A potent platelet aggregation inhibitor from venom of the viper Echis carinatus","volume":"263","author":"Gan Z.\u2010R.","year":"1988","journal-title":"J. Biol. Chem."},{"key":"e_1_2_1_14_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.86.11.4022"},{"key":"e_1_2_1_15_1","doi-asserted-by":"publisher","DOI":"10.3181\/00379727-195-43129B"},{"key":"e_1_2_1_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/pro.5560021017"},{"key":"e_1_2_1_17_1","first-page":"4925","article-title":"The basic trypsin inhibitor of bovine pancreas. Reduction with borohydride of disulfide bond linking 1\/2\u2010Cys residues 14 and 38","volume":"242","author":"Kress L.F.","year":"1967","journal-title":"J. Biol. Chem."},{"key":"e_1_2_1_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/309030a0"},{"key":"e_1_2_1_19_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1991.tb16380.x"},{"key":"e_1_2_1_20_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00244a003"}],"container-title":["Protein Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fpro.5560021018","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.5560021018","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,26]],"date-time":"2023-10-26T01:26:45Z","timestamp":1698283605000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/pro.5560021018"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1993,10]]},"references-count":19,"journal-issue":{"issue":"10","published-print":{"date-parts":[[1993,10]]}},"alternative-id":["10.1002\/pro.5560021018"],"URL":"https:\/\/doi.org\/10.1002\/pro.5560021018","archive":["Portico"],"relation":{},"ISSN":["0961-8368","1469-896X"],"issn-type":[{"value":"0961-8368","type":"print"},{"value":"1469-896X","type":"electronic"}],"subject":[],"published":{"date-parts":[[1993,10]]}}}