{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,23]],"date-time":"2025-10-23T11:01:03Z","timestamp":1761217263721},"reference-count":39,"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":4109,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[1997,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>We have studied the conformation as well as V8 protease\u2010mediated synthesis of peptide fragments, namely amino acid residues 295\u2010316 (TC\u2010peptide) of thermolysin and residues 1\u201020 (S\u2010peptide) of ribonuclease A, to examine whether \u201cconformational trapping\u201d of the product can facilitate reverse proteolysis. The circular dichroism study showed cosolvent\u2010mediated cooperative helix formation in TC\u2010peptide with attainment of about 30\u201035% helicity in the presence of 40% 1\u2010propanol and 2\u2010propanol solutions at pH 6 and 4\u00b0C. The thermal melting profiles of TC\u2010peptide in the above cosolvents were very similar. V8 protease catalyzed the synthesis of TC\u2010peptide from a 1:1 mixture of the noninteracting complementary fragments (TC295\u2010302 and TC303\u2010316) in the presence of the above cosolvents at pH 6 and 4\u00b0C. In contrast, V8 protease did not catalyze the ligation of S1\u20109 and S10\u201020, although S\u2010peptide could assume helical conformation in the presence of the cosolvent used for the semisynthetic reaction. V8 protease was able to synthesize an analog of S\u2010peptide (SA\u2010peptide) in which residues 10\u201014 were substituted (RQHMD \u2192 VAAAK). While S\u2010peptide exhibited helical conformation in the presence of aqueous propanol solutions, SA\u2010peptide displayed predominantly \u03b2\u2010sheet conformation. SA\u2010peptide showed enhanced resistance to proteolysis as compared with S\u2010peptide. Thus, failure of semisynthesis of S\u2010peptide may be a consequence of high flexibility around the 9\u201010 peptide bond due to its proximity to the helix stop signal. The results suggest that protease\u2010mediated ligations may be achieved by design and manipulation of the conformational aspects of the product.<\/jats:p>","DOI":"10.1002\/pro.5560061018","type":"journal-article","created":{"date-parts":[[2009,2,10]],"date-time":"2009-02-10T04:52:21Z","timestamp":1234241541000},"page":"2233-2241","source":"Crossref","is-referenced-by-count":8,"title":["Conformationally driven protease\u2010catalyzed splicing of peptide segments: V8 protease\u2010mediated synthesis of fragments derived from thermolysin and ribonuclease A"],"prefix":"10.1002","volume":"6","author":[{"given":"Sangaralingam","family":"Kumaran","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Debjani","family":"Datta","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rajendra 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