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Crystallization and structure determination of DAO1 from Arabidopsis thaliana : 애기장대 유래 DAO1의 결정화 및 구조 규명
DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | 이상기 | - |
dc.contributor.author | 진소희 | - |
dc.date.accessioned | 2018-12-03T01:39:19Z | - |
dc.date.available | 2021-09-23T05:26:38Z | - |
dc.date.issued | 2018-08 | - |
dc.identifier.other | 000000152144 | - |
dc.identifier.uri | https://hdl.handle.net/10371/143737 | - |
dc.description | 학위논문 (석사)-- 서울대학교 대학원 : 농업생명과학대학 농생명공학부, 2018. 8. 이상기. | - |
dc.description.abstract | Indole-3-acetic acid (IAA), the major form of the plant hormone auxin, regulates almost every aspect of plant growth and development. Therefore, auxin homeostasis is an essential issue in plants. Various pathways of synthesis, transport, conjugation, and catabolism are involved in auxin homeostasis, but its catabolic pathway has remained elusive until recent studies elucidated the presence of DIOXYGENASE FOR AUXIN OXIDATION (DAO) from Oryza sativa and Arabidopsis thaliana. DAO, a member of the 2-oxoglutarate/Fe(II)-dependent oxygenase family, constitutes a major enzyme for IAA catabolism. It catalyzes, with the cosubstrate 2-oxoglutarate, the conversion of IAA into 2-oxoindole-3-acetic acid, a functionally inactive oxidative product of IAA. Here, I report a crystal structure of the unliganded DAO1 from A. thaliana (AtDAO1) and its complex with 2-oxoglutarate. AtDAO1 is structurally homologous with members of the 2-oxoglutarate/Fe(II)-dependent oxygenase family but exhibits unique features in the prime substrate-binding site. Using liquid chromatography-tandem mass spectrometry analyses of the reaction products from various mutant enzymes, I provide structural insights into a putative binding site for the prime substrate IAA, thereby suggesting possible structural determinants for the substrate specificity of AtDAO1 toward IAA. | - |
dc.description.tableofcontents | Introduction 1
Materials and Methods 6 Cloning and purification of AtDAO1 and its mutants 6 Crystallization and data collection 15 Structure determination and refinement 18 In vitro activity assay using LC-MS/MS 20 Results Overall structure of an unliganded AtDAO1* 21 Active site of AtDOA1*. 28 Putative binding site for IAA in AtDOA1. 32 In vitro activity assay of AtDAO1 and its mutants.. 36 Discussion 41 Reference 42 Abstract in Korean 44 | - |
dc.format | application/pdf | - |
dc.format.medium | application/pdf | - |
dc.language.iso | en | - |
dc.publisher | 서울대학교 대학원 | - |
dc.subject.ddc | 630 | - |
dc.title | Crystallization and structure determination of DAO1 from Arabidopsis thaliana | - |
dc.title.alternative | 애기장대 유래 DAO1의 결정화 및 구조 규명 | - |
dc.type | Thesis | - |
dc.description.degree | Master | - |
dc.contributor.affiliation | 농업생명과학대학 농생명공학부 | - |
dc.date.awarded | 2018-08 | - |
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