DC Field | Value | Language |
---|---|---|
dc.contributor.author | Sung-Woo Kim | ko |
dc.contributor.author | Jae-Bum Kim | ko |
dc.contributor.author | Weon Sup Lee | ko |
dc.contributor.author | Woo-Hyuk Jung | ko |
dc.contributor.author | Ji-Myung Ryu | ko |
dc.contributor.author | Hyung-Wook Jang | ko |
dc.contributor.author | Young-Bae Jo | ko |
dc.contributor.author | Joon-Ki Jung | ko |
dc.contributor.author | Kim, Jung Hoe | ko |
dc.date.accessioned | 2009-12-04T08:21:05Z | - |
dc.date.available | 2009-12-04T08:21:05Z | - |
dc.date.created | 2012-02-06 | - |
dc.date.created | 2012-02-06 | - |
dc.date.issued | 2007-09 | - |
dc.identifier.citation | PROTEIN EXPRESSION AND PURIFICATION, v.55, pp.159 - 165 | - |
dc.identifier.issn | 1046-5928 | - |
dc.identifier.uri | http://hdl.handle.net/10203/14205 | - |
dc.description.abstract | Human interleukin-2 (hIL-2) was produced as a recombinant fusion protein (G3-IL-2/HF) consisting of three tandem-arranged human glucagon molecules (G3) and hIL-2. For the recovery of hIL-2, a factor Xa (FXa) cleavage sequence was introduced next to the N-terminus of hIL-2. Cleavage efficiency on this recombinant protein construct was very low because its recognition sequence was sterically hindered within the G3-IL-2/HF molecule and hence FXa access to the cleavage site was insufficient. We therefore introduced various synthetic oligopeptides upstream from the FXa cleavage site as a means to change substrate conformation and thereby increase cleavage efficiency. Among these oligopeptides, acidic or nucleophilic constructs were the most effective for the FXa-mediated cleavage of the fusion protein. In addition, insertion of various oligopeptides into the G3-IL-2/HF molecule varied the solubility of each construct depending on their physical properties. Consequently, the G3.IL-2/DF construct showed the highest final hIL-2 yields via FXa-mediated removal of the fusion partner. Lastly, we confirmed that cleavage efficiency was greatly increased but native hIL-2 was cleaved internally by non-specific cleavage when the acidic oligopeptide D4 (DDDD) was introduced upstream of the EK cleavage site within G3.IL-2/HE molecule. The G3.IL-2/HE molecule was shown to be an inefficient substrate to EK in a previous report (Biotechnol. Bioprocess Eng. (2000) 5, 13-16). (C) 2007 Elsevier Inc. All rights reserved. | - |
dc.description.sponsorship | This works was supported financially by the Ministry of Science and Technology (MOST), Republic of Korea. | en |
dc.language | English | - |
dc.language.iso | en_US | en |
dc.publisher | ACADEMIC PRESS INC ELSEVIER SCIENCE | - |
dc.subject | MALTOSE-BINDING-PROTEIN | - |
dc.subject | RECOMBINANT HUMAN GLUCAGON | - |
dc.subject | FACTOR-XA CLEAVAGE | - |
dc.subject | ESCHERICHIA-COLI | - |
dc.subject | FUSION PROTEINS | - |
dc.subject | HUMAN FERRITIN | - |
dc.subject | EXPRESSION | - |
dc.subject | PURIFICATION | - |
dc.subject | THROMBIN | - |
dc.subject | RECEPTOR | - |
dc.title | Enhanced protease cleavage efficiency on the glucagon-fused interleukin-2 by the addition of synthetic oligopeptides | - |
dc.type | Article | - |
dc.identifier.wosid | 000249631800018 | - |
dc.identifier.scopusid | 2-s2.0-34547894693 | - |
dc.type.rims | ART | - |
dc.citation.volume | 55 | - |
dc.citation.beginningpage | 159 | - |
dc.citation.endingpage | 165 | - |
dc.citation.publicationname | PROTEIN EXPRESSION AND PURIFICATION | - |
dc.identifier.doi | 10.1016/j.pep.2007.04.003 | - |
dc.embargo.liftdate | 9999-12-31 | - |
dc.embargo.terms | 9999-12-31 | - |
dc.contributor.localauthor | Kim, Jung Hoe | - |
dc.contributor.nonIdAuthor | Sung-Woo Kim | - |
dc.contributor.nonIdAuthor | Jae-Bum Kim | - |
dc.contributor.nonIdAuthor | Weon Sup Lee | - |
dc.contributor.nonIdAuthor | Woo-Hyuk Jung | - |
dc.contributor.nonIdAuthor | Ji-Myung Ryu | - |
dc.contributor.nonIdAuthor | Hyung-Wook Jang | - |
dc.contributor.nonIdAuthor | Young-Bae Jo | - |
dc.contributor.nonIdAuthor | Joon-Ki Jung | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | enterokinase | - |
dc.subject.keywordAuthor | interleukin-2 | - |
dc.subject.keywordAuthor | factor Xa | - |
dc.subject.keywordAuthor | cleavage efficiency | - |
dc.subject.keywordAuthor | fusion protein | - |
dc.subject.keywordPlus | MALTOSE-BINDING-PROTEIN | - |
dc.subject.keywordPlus | RECOMBINANT HUMAN GLUCAGON | - |
dc.subject.keywordPlus | FACTOR-XA CLEAVAGE | - |
dc.subject.keywordPlus | ESCHERICHIA-COLI | - |
dc.subject.keywordPlus | FUSION PROTEINS | - |
dc.subject.keywordPlus | HUMAN FERRITIN | - |
dc.subject.keywordPlus | EXPRESSION | - |
dc.subject.keywordPlus | PURIFICATION | - |
dc.subject.keywordPlus | THROMBIN | - |
dc.subject.keywordPlus | RECEPTOR | - |
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