DC Field | Value | Language |
---|---|---|
dc.contributor.author | Lee, NK | - |
dc.contributor.author | Koh, HR | - |
dc.contributor.author | Han, KY | - |
dc.contributor.author | Kim, SK | - |
dc.date.accessioned | 2016-04-01T08:38:08Z | - |
dc.date.available | 2016-04-01T08:38:08Z | - |
dc.date.created | 2009-08-24 | - |
dc.date.issued | 2007-12-19 | - |
dc.identifier.issn | 0002-7863 | - |
dc.identifier.other | 2008-OAK-0000018166 | - |
dc.identifier.uri | https://oasis.postech.ac.kr/handle/2014.oak/28421 | - |
dc.description.abstract | The folding of 8-17 deoxyribozyme was investigated by three-color alternating-laser excitation (3c-ALEX), a new single-molecule fluorescence resonance energy transfer (FRET) method we recently developed. Since 3c-ALEX has the capability of simultaneously sorting fluorescent molecules based on their labeling status and monitoring three interprobe distances of a biomolecule by employing three-color FRET, it is an ideal tool to study folding of multibranched molecules. The 8-17 deoxyribozyme, a DNA enzyme that cleaves a specific RNA substrate, is a good model system for a multibranched molecule, since it. has the structure of a three-way DNA junction with a bulge. Labeling all three branches of the 8-17 with different fluorescent probes, we studied its [Mg2+]-dependent folding in a Na+ buffer solution. With the stoichiometric sorting capability of 3c-ALEX, we first selected only the triply labeled 8-17 in a solution of all heterogeneous mixtures and then simultaneously measured all three interprobe distances of the selected species. Our results show that the 8-17 folds into a pyramidal form upon increasing [Mg2+]], in a similar way with [Zn2+] as found in an earlier study conducted at the ensemble level. The apparent dissociation constant of Mg2+ was more than 100 times larger than that of Zn2+ and showed considerable variance with buffer concentration. No clear sign of two-step folding was observed for Mg2+, in contrast to the case of Zn2+. Compared with the hammerhead ribozyme, the 8-17 was found to require 10 times higher [Mg2+] to undergo folding. By comparison with the folding of several inactive 8-17 analogues, we found that the two conserved sequences (A and G) of the triad loop of the shortest branch are critical elements for folding, especially for the folding at low [Mg2+]. Our results suggest that the role of the stem loop is to provide a scaffold for the two bases to be properly positioned for the necessary interaction and that the two bases are directly involved in the interaction that plays a critical role in folding. This work demonstrates that 3c-ALEX is a powerful single-molecule method to study the structure and folding of complex and multibranched biomolecules. | - |
dc.description.statementofresponsibility | X | - |
dc.language | English | - |
dc.publisher | AMER CHEMICAL SOC | - |
dc.relation.isPartOf | JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | - |
dc.subject | RNA-CLEAVING DEOXYRIBOZYME | - |
dc.subject | RESONANCE ENERGY-TRANSFER | - |
dc.subject | IN-VITRO SELECTION | - |
dc.subject | DNA ENZYME | - |
dc.subject | HAMMERHEAD RIBOZYME | - |
dc.subject | LOGIC GATES | - |
dc.subject | ION-BINDING | - |
dc.subject | FRET | - |
dc.subject | DNAZYME | - |
dc.subject | SPECTROSCOPY | - |
dc.title | FOLDING OF 8-17 DEOXYRIBOZYME STUDIED BY THREE-COLOR ALTERNATING-LASER EXCITATION OF SINGLE MOLECULES | - |
dc.type | Article | - |
dc.contributor.college | 시스템생명공학부 | - |
dc.identifier.doi | 10.1021/JA0725145 | - |
dc.author.google | Lee, NK | - |
dc.author.google | Koh, HR | - |
dc.author.google | Han, KY | - |
dc.author.google | Kim, SK | - |
dc.relation.volume | 129 | - |
dc.relation.issue | 50 | - |
dc.relation.startpage | 15526 | - |
dc.relation.lastpage | 15534 | - |
dc.contributor.id | 10206847 | - |
dc.relation.journal | JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | - |
dc.relation.index | SCI급, SCOPUS 등재논문 | - |
dc.relation.sci | SCI | - |
dc.collections.name | Journal Papers | - |
dc.type.rims | ART | - |
dc.identifier.bibliographicCitation | JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, v.129, no.50, pp.15526 - 15534 | - |
dc.identifier.wosid | 000251581900036 | - |
dc.date.tcdate | 2019-02-01 | - |
dc.citation.endPage | 15534 | - |
dc.citation.number | 50 | - |
dc.citation.startPage | 15526 | - |
dc.citation.title | JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | - |
dc.citation.volume | 129 | - |
dc.contributor.affiliatedAuthor | Lee, NK | - |
dc.identifier.scopusid | 2-s2.0-37849026396 | - |
dc.description.journalClass | 1 | - |
dc.description.journalClass | 1 | - |
dc.description.wostc | 34 | - |
dc.description.scptc | 17 | * |
dc.date.scptcdate | 2018-05-121 | * |
dc.type.docType | Article | - |
dc.subject.keywordPlus | IN-VITRO SELECTION | - |
dc.subject.keywordPlus | DNA ENZYME | - |
dc.subject.keywordPlus | HAMMERHEAD RIBOZYME | - |
dc.subject.keywordPlus | 8-17 DEOXYRIBOZYME | - |
dc.subject.keywordPlus | ION-BINDING | - |
dc.subject.keywordPlus | RNA | - |
dc.subject.keywordPlus | FRET | - |
dc.subject.keywordPlus | SPECTROSCOPY | - |
dc.subject.keywordPlus | CLEAVAGE | - |
dc.subject.keywordPlus | BIOSENSOR | - |
dc.relation.journalWebOfScienceCategory | Chemistry, Multidisciplinary | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Chemistry | - |
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