{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T22:23:24Z","timestamp":1777501404019,"version":"3.51.4"},"reference-count":25,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2001,4,19]],"date-time":"2001-04-19T00:00:00Z","timestamp":987638400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Proteins"],"published-print":{"date-parts":[[2001,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Rhodostomin (Rho) is a snake venom protein isolated from <jats:italic>Calloselasma rhodostoma<\/jats:italic>. Rho is a disintegrin that inhibits platelet aggregation by blocking the binding of fibrinogen to the integrin \u03b1<jats:sub>IIb<\/jats:sub>\u03b2<jats:sub>3<\/jats:sub> of platelets. Rho produced in <jats:italic>Escherichia coli<\/jats:italic> inhibited platelet aggregation with a <jats:italic>K<\/jats:italic><jats:sub>I<\/jats:sub> value of 263 nM. Although functional, Rho produced in <jats:italic>E. coli<\/jats:italic> is misfolded based on our 2D and 3D NMR studies. In order to correct the folding problem, Rho was expressed in <jats:italic>Pichia pastoris<\/jats:italic>. The recombinant Rho expressed in <jats:italic>P. pastoris<\/jats:italic> inhibited platelet aggregation with a resulting <jats:italic>K<\/jats:italic><jats:sub>I<\/jats:sub> value of 70 nM. This is the same potency as that of native Rho. CD analysis showed that the secondary structures of Rho are pH\u2010independent and contain 3.5\u20137.9% \u03b1\u2010helix, 48.2\u201350.5% \u03b2\u2010structures, and 42.3\u201347% coil. The sequential assignment and structure analysis of Rho were obtained using 2D and 3D <jats:sup>15<\/jats:sup>N\u2010edited NMR spectra. These results provide the first direct evidence that highly disulfide\u2010bonded disintegrin can be expressed in <jats:italic>P. pastoris<\/jats:italic> with the correct fold. This evidence may serve as the basis for exploring the structure and function relationships as well as the dynamics of disintegrin and its variants. Proteins 2001;43:499\u2013508. \u00a9 2001 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/prot.1061","type":"journal-article","created":{"date-parts":[[2002,8,25]],"date-time":"2002-08-25T04:30:39Z","timestamp":1030249839000},"page":"499-508","source":"Crossref","is-referenced-by-count":36,"title":["Expression in <i>Pichia pastoris<\/i> and characterization by circular dichroism and NMR of rhodostomin"],"prefix":"10.1002","volume":"43","author":[{"given":"Rey\u2010Ting","family":"Guo","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Li\u2010Jer","family":"Chou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yen\u2010Chin","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chiu\u2010Yueh","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Koteppa","family":"Pari","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chauying J.","family":"Jen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Szecheng J.","family":"Lo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shou\u2010Ling","family":"Huang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chun\u2010Yann","family":"Lee","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tsai\u2010Wang","family":"Chang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Woei\u2010Jer","family":"Chaung","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2001,4,19]]},"reference":[{"key":"e_1_2_8_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/0955-0674(92)90098-W"},{"key":"e_1_2_8_3_2","first-page":"289","article-title":"Disintegrins and other naturally occurring antagonists of platelet fibrinogen receptors","volume":"31","author":"Niewiarowski S","year":"1994","journal-title":"Semin Hematol"},{"key":"e_1_2_8_4_2","doi-asserted-by":"publisher","DOI":"10.3181\/00379727-219-44322"},{"key":"e_1_2_8_5_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1862345"},{"key":"e_1_2_8_6_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00244a003"},{"key":"e_1_2_8_7_2","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1993.1439"},{"key":"e_1_2_8_8_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1399-3011.1996.tb00835.x"},{"key":"e_1_2_8_9_2","doi-asserted-by":"crossref","first-page":"9359","DOI":"10.1016\/S0021-9258(18)92826-7","article-title":"Barbourin. 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