{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/84812"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/84812","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Biochemical and Structural Studies of Pseudouridine 55 Synthase","abstract":"Sequence alignment of the families of PsiS identifies the conserved aspartic acid discussed previously as the strictly conserved amino acid. Structural alignment identifies the conserved tyrosine, which is part of hydrophobic core in the active site. The role of the conserved tyrosine is less obvious in addition to apparent structural role. Enzymatic activity assay reveals that mutating T. maritima Tyr67 to any other amino acids abolishes the enzymatic activity. Structure of T. maritima Y67F in complex with the 5FU-RNA reveals, however, that the same 5FhPsi product formed by wild-type Psi55S is also found in the active site. Furthermore, HPLC analysis indicates that 5FhPsi is also formed when 5FU RNA was incubated with either E. coli Y76F or Y76L but not with Y76A. The combined information from structural, biochemical and mutational studies allows us to propose the likely role of the conserved tyrosine as a general base for proton abstraction in PsiS-catalyzed reaction as well as structural role of the hydrophobic phenol ring in the active site.","abstract_html":"Sequence alignment of the families of PsiS identifies the conserved aspartic acid discussed previously as the strictly conserved amino acid. Structural alignment identifies the conserved tyrosine, which is part of hydrophobic core in the active site. The role of the conserved tyrosine is less obvious in addition to apparent structural role. Enzymatic activity assay reveals that mutating T. maritima Tyr67 to any other amino acids abolishes the enzymatic activity. Structure of T. maritima Y67F in complex with the 5FU-RNA reveals, however, that the same 5FhPsi product formed by wild-type Psi55S is also found in the active site. Furthermore, HPLC analysis indicates that 5FhPsi is also formed when 5FU RNA was incubated with either E. coli Y76F or Y76L but not with Y76A. The combined information from structural, biochemical and mutational studies allows us to propose the likely role of the conserved tyrosine as a general base for proton abstraction in PsiS-catalyzed reaction as well as structural role of the hydrophobic phenol ring in the active site.","abstract_has_math":false,"creators":["Phannachet, Kulwadee"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Biochemistry","degree_department":null,"school":null,"contributors":["Huang, Raven H."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2015,"date_issued":"2015-09-25T22:28:02Z","date_published":"2015-09-25T22:28:02Z","updated_at":"2026-07-22T22:26:24Z","subjects":["Chemistry, Biochemistry"],"languages":["eng"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["(MiAaPQ)AAI3182354"],"render_values":[{"text":"(MiAaPQ)AAI3182354","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/84812","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Huang, Raven H."]},{"key":"dc:creator","label":"Author","values":["Phannachet, Kulwadee"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2015-09-25T22:28:02Z","10000-01-01","2005"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biochemistry"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Chemistry, Biochemistry"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/84812","(MiAaPQ)AAI3182354"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Sequence alignment of the families of PsiS identifies the conserved aspartic acid discussed previously as the strictly conserved amino acid. Structural alignment identifies the conserved tyrosine, which is part of hydrophobic core in the active site. The role of the conserved tyrosine is less obvious in addition to apparent structural role. Enzymatic activity assay reveals that mutating T. maritima Tyr67 to any other amino acids abolishes the enzymatic activity. Structure of T. maritima Y67F in complex with the 5FU-RNA reveals, however, that the same 5FhPsi product formed by wild-type Psi55S is also found in the active site. Furthermore, HPLC analysis indicates that 5FhPsi is also formed when 5FU RNA was incubated with either E. coli Y76F or Y76L but not with Y76A. The combined information from structural, biochemical and mutational studies allows us to propose the likely role of the conserved tyrosine as a general base for proton abstraction in PsiS-catalyzed reaction as well as structural role of the hydrophobic phenol ring in the active site.","Made available in DSpace on 2015-09-25T22:28:02Z (GMT). No. of bitstreams: 2 license.txt: 4848 bytes, checksum: 96035ab3f5e1c23cc7138a224ce498bd (MD5) 3182354.pdf: 5925328 bytes, checksum: 5b212aaec6eb55b7be3251c0a69aabc7 (MD5) Previous issue date: 2005","Embargo set by: Seth Robbins for item 86093 Lift date: Forever Reason: Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","U of I Only","118 p.","Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 2005."]},{"key":"dc:title","label":"Title","values":["Biochemical and Structural Studies of Pseudouridine 55 Synthase"]}]}],"canonical_facts":{"dc:contributor":["Huang, Raven H."],"dc:creator":["Phannachet, Kulwadee"],"dc:date":["2015-09-25T22:28:02Z","10000-01-01","2005"],"dc:description":["Sequence alignment of the families of PsiS identifies the conserved aspartic acid discussed previously as the strictly conserved amino acid. Structural alignment identifies the conserved tyrosine, which is part of hydrophobic core in the active site. The role of the conserved tyrosine is less obvious in addition to apparent structural role. Enzymatic activity assay reveals that mutating T. maritima Tyr67 to any other amino acids abolishes the enzymatic activity. Structure of T. maritima Y67F in complex with the 5FU-RNA reveals, however, that the same 5FhPsi product formed by wild-type Psi55S is also found in the active site. Furthermore, HPLC analysis indicates that 5FhPsi is also formed when 5FU RNA was incubated with either E. coli Y76F or Y76L but not with Y76A. The combined information from structural, biochemical and mutational studies allows us to propose the likely role of the conserved tyrosine as a general base for proton abstraction in PsiS-catalyzed reaction as well as structural role of the hydrophobic phenol ring in the active site.","Made available in DSpace on 2015-09-25T22:28:02Z (GMT). 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