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dc.contributor.advisorHenzl, Michael T.eng
dc.contributor.authorNdubuka, Kellyeng
dc.date.issued2011eng
dc.date.submitted2011 Springeng
dc.descriptionTitle from PDF of title page (University of Missouri--Columbia, viewed on June 13, 2011).eng
dc.descriptionThe entire thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file; a non-technical public abstract appears in the public.pdf file.eng
dc.descriptionThesis advisor: Dr. Michael T. Henzl.eng
dc.descriptionIncludes bibliographical references.eng
dc.descriptionM.S. University of Missouri--Columbia 2011.eng
dc.description.abstractParvalbumins (PVs) are vertebrate-specific proteins (Mr 12,000), which harbor two EF-hand motifs known as the CD and EF sites. Although the CD and EF sites are typically high-affinity sites, the mammalian [beta]-PV exhibits highly attenuated divalent ion-affinity. The physical basis for this attenuation remains unclear. A clarification of this behavior could advance our understanding of EF-hand protein structure-affinity relationships. The question arises as to whether the difference in divalent ion-binding affinity in these proteins derives from local differences in and around the immediate binding site, or whether remote structural determinants play a role. To address this matter, site-directed mutagenesis was performed on rat [beta]-PV at positions 49, 50, 57, 58, 59, and 60, making it identical to CPV3 at 27 of 30 residues at the CD site. Divalent ion affinity and thermal stability were evaluated for each of the variants using isothermal titration calorimetry and differential scanning calorimetry, respectively. The mutations resulted in an increase in melting temperature. However, the increases in the Ca2+-free state indicating heightened stability were small in comparison to CPV3. These findings suggest that structural determinants outside the metal ion-binding motif significantly affect the attenuated binding affinity observed at the CD site in rat [beta]-PV.eng
dc.format.extentviii, 88 pageseng
dc.identifier.urihttp://hdl.handle.net/10355/11499
dc.languageEnglisheng
dc.publisherUniversity of Missouri--Columbiaeng
dc.relation.ispartofcommunityUniversity of Missouri--Columbia. Graduate School. Theses and Dissertationseng
dc.rightsOpenAccess.eng
dc.rights.licenseThis work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License.
dc.sourceSubmitted by University of Missouri--Columbia Graduate School.eng
dc.subject.lcshAlbumins -- Structure-activity relationshipseng
dc.subject.lcshVolumetric analysiseng
dc.subject.lcshCalorimetryeng
dc.subject.lcshCalmodulineng
dc.subject.lcshStructure-activity relationships (Biochemistry)eng
dc.subject.lcshAmino acids -- Effect of heat oneng
dc.subject.lcshTemperature -- Physiological effecteng
dc.subject.lcshRats -- Physiology -- Experimentseng
dc.titleDivalent ion-binding and thermal stability studies on rat [beta]-parvalbumin and the evidence of influential distant amino acid residues affecting CD site ion affinityeng
dc.typeThesiseng
thesis.degree.disciplineBiochemistry (MU)eng
thesis.degree.grantorUniversity of Missouri--Columbiaeng
thesis.degree.levelMasterseng
thesis.degree.nameM.S.eng


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