Characterization of ubiquitin C-terminal hydrolase 1 (YUH1) from Saccharomyces cerevisiae expressed in recombinant Escherichia coli
DC Field | Value | Language |
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dc.contributor.author | Yu, Hyun-Ah | - |
dc.contributor.author | Kim, Sung-Gun | - |
dc.contributor.author | Kim, Eun-Jeong | - |
dc.contributor.author | Lee, Woo-Jong | - |
dc.contributor.author | Kim, Dae-Ok | - |
dc.contributor.author | Park, Kyungmoon | - |
dc.contributor.author | Park, Yong-Cheol | - |
dc.contributor.author | Seo, Jin-Ho | - |
dc.date.accessioned | 2022-01-14T07:41:29Z | - |
dc.date.available | 2022-01-14T07:41:29Z | - |
dc.date.created | 2022-01-14 | - |
dc.date.issued | 2007-11 | - |
dc.identifier.issn | 1046-5928 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/hongik/handle/2020.sw.hongik/23523 | - |
dc.description.abstract | The YUH1 gene coding for ubiquitin C-terminal hydrolase 1, a deubiquitinating enzyme, was cloned from the Saccharomyces cerevisiae genomic DNA and expressed in Escherichia coh. YUH1 was fused with the 6 histidine tag at the N-terminus (H6YUH1) or C-terminus (YUH1H6) and purified by an immobilized metal affinity chromatography with high purity. By using a fluorogenic substrate, Z-Arg-Leu-Arg-Gly-Gly-AMC, the deubiquitinating activities for H6YUH1 (1.72 U/mg) and YUH1H6 (1.61 U/mg) were about 18 times higher than 0.092 U/mg for H6UBP1, ubiquitin specific protease 1 of S. cerevisiae containing the 6 histidine residue at the N-terminus which is normally used in protein engineering. YUH1 had the optimal temperature of 27 degrees C and acidity of pH 8.5. Analysis of thermal deactivation kinetics of H6YUH1 estimated 3.2 and 1.4 h of half lives at 4 and 52 degrees C, respectively. Immobilization onto the Ni-NTA affinity resin and environmental modulation were carried out to improve the stability of YUH1. Incubation of the immobilized YUH1 in 50% glycerol solution at -20 degrees C resulted in 52 degrees A of decrease in specific activity for 7 days, corresponding to a 2.7-fold increase compared with that of the free YUH1 incubated in the same solution at 4 degrees C. (c) 2007 Elsevier Inc. All rights reserved. | - |
dc.language | 영어 | - |
dc.language.iso | en | - |
dc.publisher | ACADEMIC PRESS INC ELSEVIER SCIENCE | - |
dc.subject | DEUBIQUITINATING ENZYMES | - |
dc.subject | PURIFICATION | - |
dc.subject | FUSION | - |
dc.subject | PROTEASE | - |
dc.subject | CLONING | - |
dc.title | Characterization of ubiquitin C-terminal hydrolase 1 (YUH1) from Saccharomyces cerevisiae expressed in recombinant Escherichia coli | - |
dc.type | Article | - |
dc.contributor.affiliatedAuthor | Park, Kyungmoon | - |
dc.identifier.doi | 10.1016/j.pep.2007.07.005 | - |
dc.identifier.scopusid | 2-s2.0-34848887990 | - |
dc.identifier.wosid | 000250639100003 | - |
dc.identifier.bibliographicCitation | PROTEIN EXPRESSION AND PURIFICATION, v.56, no.1, pp.20 - 26 | - |
dc.relation.isPartOf | PROTEIN EXPRESSION AND PURIFICATION | - |
dc.citation.title | PROTEIN EXPRESSION AND PURIFICATION | - |
dc.citation.volume | 56 | - |
dc.citation.number | 1 | - |
dc.citation.startPage | 20 | - |
dc.citation.endPage | 26 | - |
dc.type.rims | ART | - |
dc.type.docType | Article | - |
dc.description.journalClass | 1 | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Biochemistry & Molecular Biology | - |
dc.relation.journalResearchArea | Biotechnology & Applied Microbiology | - |
dc.relation.journalWebOfScienceCategory | Biochemical Research Methods | - |
dc.relation.journalWebOfScienceCategory | Biochemistry & Molecular Biology | - |
dc.relation.journalWebOfScienceCategory | Biotechnology & Applied Microbiology | - |
dc.subject.keywordPlus | DEUBIQUITINATING ENZYMES | - |
dc.subject.keywordPlus | PURIFICATION | - |
dc.subject.keywordPlus | FUSION | - |
dc.subject.keywordPlus | PROTEASE | - |
dc.subject.keywordPlus | CLONING | - |
dc.subject.keywordAuthor | YUH1 | - |
dc.subject.keywordAuthor | ubiquitin c-terminal hydrolase | - |
dc.subject.keywordAuthor | Saccharomyces cerevisiae | - |
dc.subject.keywordAuthor | recombinant Escherichia coli | - |
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