Succinate-binding site of succinyl-CoA synthetase

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atmire.migration.oldid2785
dc.contributor.advisorFraser, Marie
dc.contributor.authorHuang, Ji
dc.date.accessioned2014-12-15T23:20:16Z
dc.date.available2015-02-23T08:00:32Z
dc.date.issued2014-12-15
dc.date.submitted2014en
dc.description.abstractSuccinyl-CoA synthetase (SCS) is an essential enzyme that catalyzes substrate-level phosphorylation in the citric acid cycle. In my study, the pig GTP-specific SCS (GTPSCS) was overproduced in E. coli BL21(DE3), and purified through three different columns: Ni-NTA affinity, Superdex-200 prep grade size exclusion and Hitrap Blue High Performance affinity. A crystal of GTPSCS in complex with coenzyme A (CoA) diffracted to 2.1 Å. The CoA-binding site was determined to be located in the N-terminal domain of the α-subunit of the enzyme. A crystal of GTPSCS in complex with Mg2+-succinate and CoA was grown in polyethylene glycol 3350, ammonium succinate and Hepes (pH 7.0) and diffracted to 2.2 Å. The succinate-binding site of GTPSCS was determined to be located in the C-terminal domain of the β-subunit of the enzyme. The discovery of the succinate-binding site of SCS supports the view that the succinyl phosphate complex can be formed during the catalytic reaction.en_US
dc.identifier.citationHuang, J. (2014). Succinate-binding site of succinyl-CoA synthetase (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca. doi:10.11575/PRISM/27933en_US
dc.identifier.doihttp://dx.doi.org/10.11575/PRISM/27933
dc.identifier.urihttp://hdl.handle.net/11023/1962
dc.language.isoeng
dc.publisher.facultyGraduate Studies
dc.publisher.institutionUniversity of Calgaryen
dc.publisher.placeCalgaryen
dc.rightsUniversity of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.
dc.subjectBiochemistry
dc.subject.classificationSuccinate-binding siteen_US
dc.subject.classificationGTP-specific succinyl-CoA synthetaseen_US
dc.subject.classificationCrystalen_US
dc.subject.classificationStructureen_US
dc.titleSuccinate-binding site of succinyl-CoA synthetase
dc.typemaster thesis
thesis.degree.disciplineBiological Sciences
thesis.degree.grantorUniversity of Calgary
thesis.degree.nameMaster of Science (MSc)
ucalgary.item.requestcopytrue
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