DC Field | Value | Language |
---|---|---|
dc.contributor.author | Sathya Murthy, Madhavi | ko |
dc.contributor.author | Kim, Che Lin | ko |
dc.contributor.author | Bang, You Lim | ko |
dc.contributor.author | Kim, Young Sik | ko |
dc.contributor.author | Jang, Ju Woong | ko |
dc.contributor.author | Lee, Gyun-Min | ko |
dc.date.accessioned | 2015-04-08T04:50:01Z | - |
dc.date.available | 2015-04-08T04:50:01Z | - |
dc.date.created | 2014-12-12 | - |
dc.date.created | 2014-12-12 | - |
dc.date.created | 2014-12-12 | - |
dc.date.issued | 2015-03 | - |
dc.identifier.citation | BIOTECHNOLOGY AND BIOENGINEERING, v.112, no.3, pp.560 - 568 | - |
dc.identifier.issn | 0006-3592 | - |
dc.identifier.uri | http://hdl.handle.net/10203/195589 | - |
dc.description.abstract | Bone morphogenetic protein-7 (BMP-7) is synthesized as a precursor that requires proteolytic cleavage of the propeptide by proprotein convertases (PCs) for its functional activity. A high-level expression of BMP-7 in CHO cells (CHO-BMP-7) resulted in secretion of a mixture of inactive precursor and active BMP-7. In an effort to achieve efficient processing of BMP-7 in CHO cells, PCs responsible for cleavage of the precursors in CHO cells were characterized. Analysis of the mRNA expression levels of four PCs (furin, PACE4, PC5/6, and PC7) revealed that only furin and PC7 genes are expressed in CHO-BMP-7 cells. Specific inhibition of the PCs by hexa-D-arginine (D6R) or decanoyl-RVKR-chloromethyl ketone (RVKR-CMK) further revealed that furin is mainly responsible for the proteolytic processing of BMP-7. To identify a more efficient PC for BMP-7 processing, the four PC genes were transiently expressed in CHO-BMP-7 cells, respectively. Among these, PC5/6 was found to be the most efficient in BMP-7 processing. Stable overexpression of PC5/6C, a secreted form of PC5/6, significantly improved mature BMP-7 production in CHO-BMP-7 cells. When the maximum BMP-7 concentration was obtained in the culture of CHO-BMP-7 cells, approximately 88% of BMP-7 was unprocessed. In contrast, no precursor was found in the culture of PC5/6C-overexpressing cells (clone #97). Furthermore, the in vitro biological activity of the mature BMP-7 from PC5/6C-overexpressing cells was comparable to that from CHO-BMP-7 cells. Taken together, the present results indicate that overexpression of PC5/6C in CHO-BMP-7 cells is an efficient means of increasing the yield of BMP-7. Biotechnol. Bioeng. 2015;112: 560-568. | - |
dc.language | English | - |
dc.publisher | WILEY-BLACKWELL | - |
dc.title | Characterization and Expression of Proprotein Convertases in CHO Cells: Efficient Proteolytic Maturation of Human Bone Morphogenetic Protein-7 | - |
dc.type | Article | - |
dc.identifier.wosid | 000349161400013 | - |
dc.identifier.scopusid | 2-s2.0-84922808712 | - |
dc.type.rims | ART | - |
dc.citation.volume | 112 | - |
dc.citation.issue | 3 | - |
dc.citation.beginningpage | 560 | - |
dc.citation.endingpage | 568 | - |
dc.citation.publicationname | BIOTECHNOLOGY AND BIOENGINEERING | - |
dc.identifier.doi | 10.1002/bit.25458 | - |
dc.contributor.localauthor | Lee, Gyun-Min | - |
dc.contributor.nonIdAuthor | Bang, You Lim | - |
dc.contributor.nonIdAuthor | Kim, Young Sik | - |
dc.contributor.nonIdAuthor | Jang, Ju Woong | - |
dc.description.isOpenAccess | N | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | proprotein convertase | - |
dc.subject.keywordAuthor | BMP-7 | - |
dc.subject.keywordAuthor | CHO cells | - |
dc.subject.keywordAuthor | BMP-7 precursors | - |
dc.subject.keywordAuthor | PC5 | - |
dc.subject.keywordAuthor | 6 Delta C | - |
dc.subject.keywordPlus | HAMSTER OVARY CELLS | - |
dc.subject.keywordPlus | PRECURSOR CLEAVAGE | - |
dc.subject.keywordPlus | SECRETORY PATHWAY | - |
dc.subject.keywordPlus | TISSUE INHIBITORS | - |
dc.subject.keywordPlus | BREAST-CANCER | - |
dc.subject.keywordPlus | IN-VIVO | - |
dc.subject.keywordPlus | FURIN | - |
dc.subject.keywordPlus | CULTURE | - |
dc.subject.keywordPlus | PACE4 | - |
dc.subject.keywordPlus | BMP-7 | - |
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