DC Field | Value | Language |
---|---|---|
dc.contributor.author | Choi, G | - |
dc.contributor.author | Ha, NC | - |
dc.contributor.author | Kim, SW | - |
dc.contributor.author | Kim, DH | - |
dc.contributor.author | Park, S | - |
dc.contributor.author | Oh, BH | - |
dc.contributor.author | Choi, KY | - |
dc.date.accessioned | 2016-03-31T13:34:24Z | - |
dc.date.available | 2016-03-31T13:34:24Z | - |
dc.date.created | 2009-03-17 | - |
dc.date.issued | 2000-02-08 | - |
dc.identifier.issn | 0006-2960 | - |
dc.identifier.other | 2000-OAK-0000001146 | - |
dc.identifier.uri | https://oasis.postech.ac.kr/handle/2014.oak/20127 | - |
dc.description.abstract | Delta(5)-3-ketosteroid isomerase (KSI) catalyzes the allylic isomerization of Delta(5)-3-ketosteroids at a rate approaching the diffusion Limit by an intramolecular transfer of a proton. Despite the extensive studies on the catalytic mechanism, it still remains controversial whether the catalytic residue Asp-99 donates a hydrogen bond to the steroid or to Tyr-14. To clarify the role of Asp-99 in the catalysis, two single mutants of D99E and D99L and three double mutants of Y14F/D99E, Y14F/D99N, and Y14F/D99L have been prepared by site-directed mutagenesis, The D99E mutant whose side chain at position 99 is longer by an additional methylene group exhibits nearly the same k(cat) as the wild-type while the D99L mutant exhibits ca. 125-fold lower k(cat) than that of the wild-type. The mutations made at positions 14 and 99 exert synergistic or partially additive effect on k(cat) in the double mutants, which is inconsistent with the mechanism based on the hydrogen-bonded catalytic diad, Asp-99 COOH...Tyr-14 OH ... C3-O of the steroid. The crystal structure of D99E/D38N complexed with equilenin, an intermediate analogue, at 1.9 Angstrom resolution reveals that the distance between Tyr-14 O eta and Glu-99 O epsilon is Ca. 4.2 Angstrom, which is beyond the range for a hydrogen bond, and that the distance between Glu-99 O epsilon and C3-O of the steroid is maintained to be ca, 2.4 Angstrom, short enough for a hydrogen bond to be formed. Taken together, these results strongly support the idea that Asp-99 contributes to the catalysis by donating a hydrogen bond directly to the intermediate. | - |
dc.description.statementofresponsibility | X | - |
dc.language | English | - |
dc.publisher | AMER CHEMICAL SOC | - |
dc.relation.isPartOf | BIOCHEMISTRY | - |
dc.subject | 3-OXO-DELTA(5)-STEROID ISOMERASE | - |
dc.subject | DELTA-5-3-KETOSTEROID ISOMERASE | - |
dc.subject | INTERMEDIATE | - |
dc.subject | BINDING | - |
dc.subject | PROTON | - |
dc.subject | MUTANT | - |
dc.subject | D38N | - |
dc.title | Asp-99 donates a hydrogen bond not to tyr-14 but to the steroid directly in the catalytic mechanism of Delta(5)-3-ketosteroid isomerase from Pseudomonas putida biotype B | - |
dc.type | Article | - |
dc.contributor.college | 생명과학과 | - |
dc.identifier.doi | 10.1021/bi991579k | - |
dc.author.google | Choi, G | - |
dc.author.google | Ha, NC | - |
dc.author.google | Kim, SW | - |
dc.author.google | Kim, DH | - |
dc.author.google | Park, S | - |
dc.author.google | Oh, BH | - |
dc.author.google | Choi, KY | - |
dc.relation.volume | 39 | - |
dc.relation.issue | 5 | - |
dc.relation.startpage | 903 | - |
dc.relation.lastpage | 909 | - |
dc.contributor.id | 10052985 | - |
dc.relation.journal | BIOCHEMISTRY | - |
dc.relation.index | SCI급, SCOPUS 등재논문 | - |
dc.relation.sci | SCI | - |
dc.collections.name | Journal Papers | - |
dc.type.rims | ART | - |
dc.identifier.bibliographicCitation | BIOCHEMISTRY, v.39, no.5, pp.903 - 909 | - |
dc.identifier.wosid | 000085172900007 | - |
dc.date.tcdate | 2019-01-01 | - |
dc.citation.endPage | 909 | - |
dc.citation.number | 5 | - |
dc.citation.startPage | 903 | - |
dc.citation.title | BIOCHEMISTRY | - |
dc.citation.volume | 39 | - |
dc.contributor.affiliatedAuthor | Oh, BH | - |
dc.contributor.affiliatedAuthor | Choi, KY | - |
dc.identifier.scopusid | 2-s2.0-0034620560 | - |
dc.description.journalClass | 1 | - |
dc.description.journalClass | 1 | - |
dc.description.wostc | 36 | - |
dc.type.docType | Article | - |
dc.subject.keywordPlus | 3-OXO-DELTA(5)-STEROID ISOMERASE | - |
dc.subject.keywordPlus | DELTA-5-3-KETOSTEROID ISOMERASE | - |
dc.subject.keywordPlus | INTERMEDIATE | - |
dc.subject.keywordPlus | BINDING | - |
dc.subject.keywordPlus | PROTON | - |
dc.subject.keywordPlus | MUTANT | - |
dc.subject.keywordPlus | D38N | - |
dc.relation.journalWebOfScienceCategory | Biochemistry & Molecular Biology | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Biochemistry & Molecular Biology | - |
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