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Crystal structure of human endothelial overexpressed lipopolysaccharide-associated factor 1

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dc.contributor.advisor한병우-
dc.contributor.authorMinju Kim-
dc.date.accessioned2017-10-31T08:19:11Z-
dc.date.available2018-08-01-
dc.date.issued2017-08-
dc.identifier.other000000146622-
dc.identifier.urihttps://hdl.handle.net/10371/137928-
dc.description학위논문 (석사)-- 서울대학교 대학원 약학대학 약학과, 2017. 8. 한병우.-
dc.description.abstractEndothelial-overexpressed LPS-associated factor 1 (EOLA1) is a novel gene discovered in an effort to identify genes expressed in endothelial cells by activation of lipopolysaccharide. Previous study demonstrates that knocking down EOLA1 stimulated interleukin-6 (IL6) and apoptosis in the treatment of LPS in human umbilical vein endothelial cells (HUVEC). Analysis of the sequence of EOLA1, revealed that it has ASCH (activating signal cointegrator-1 homology) domain containing a unique β-barrel fold similar to pseudouridine synthase and archaeosine transglycosylase (PUA) domain. Because PUA domain is an ancient RNA binding domain, both ASCH and PUA domain appear to have arisen from a common RNA binding precursor. Here, I report the crystal structure of EOLA1 at 1.7 Å resolution by single wavelength anomalous dispersion (SAD) method using selenomethionine derivative crystals to solve the phasing problem. I demonstrated that EOLA1 could bind with RNA through gel mobility shift assay. Based on these results, I propose that EOLA1 is a RNA binding domain and would play regulatory roles in transcription or in protection of HUVEC injury with inflammation.-
dc.description.tableofcontentsI.Introduction 1
II.Materials and Methods 4
1. Materials 4
2. Methods 4
2.1. Cloning 4
2.2. Overexpression 5
2.3. Purification 5
2.4. Crystallization 6
2.5. X-ray data collection and structure determination 6
2.6. Electrophoretic mobility shift assay (EMSA) 7
III. Results and Discussion 8
1. Cloning 8
2. Overexpression and purification 8
3. Crystallization 16
4. X-ray data collection and structure determination 17
5. Electrophoretic mobility shift assay (EMSA) 19
6. Overall structure of EOLA1 21
7. Structure analysis 22
IV. References 27
V. Acknowledgement 30
Abstract in korean 31
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dc.formatapplication/pdf-
dc.format.extent1763694 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 대학원-
dc.subjectendothelial-overexpressed LPS associated factor 1-
dc.subjectRNA binding domain-
dc.subjectPUA domain-
dc.subjectcrystal structure-
dc.subjectprotein X-ray crystallography-
dc.subject.ddc615-
dc.titleCrystal structure of human endothelial overexpressed lipopolysaccharide-associated factor 1-
dc.typeThesis-
dc.description.degreeMaster-
dc.contributor.affiliation약학대학 약학과-
dc.date.awarded2017-08-
Appears in Collections:
College of Pharmacy (약학대학)Dept. of Pharmacy (약학과)Theses (Master's Degree_약학과)
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