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dc.contributor.advisorPastrana-Ríos, Belinda
dc.contributor.authorMeza-Venencia, Verónica
dc.date.accessioned2018-10-05T17:58:21Z
dc.date.available2018-10-05T17:58:21Z
dc.date.issued2005
dc.identifier.urihttps://hdl.handle.net/20.500.11801/1005
dc.description.abstractHuman centrin 2 (Hcen 2) is an acidic centrosomal protein of 19,738 Da, that is comprised of four subdomains, called EF-hands, which represent the calcium-binding sites. Human centrin 2 gene locus is codified on chromosomes X and had been designated as Hcetn2. Hcen2 may play a role in centrosome duplication and segregation during mitosis. Transformed bacterial E. coli cells with a recombinant molecule containing the Hcetn2 gene were grown in either 2xYT broth or 13C isotope enriched minimal media and induced to over-express the Hcen 2 and 13C-Hcen 2, respectively. These recombinant proteins were then purified by using hydrophobic affinity chromatography followed by several anion exchange chromatographic separations. Highly pure Human centrin 2 and 13C-homogenously labeled protein were obtained for biophysical studies. Differential scanning calorimetric (DSC) study of Hcen 2 showed that this is a very stable protein with a pre-transition temperature around 85 °C and a thermal transition temperature of 120 °C.en_US
dc.description.abstractLa centrina humana 2 (Hcen 2) es una proteína acídica del centrosoma con un peso molecular de 19, 738 Da. Está compuesta por cuatro sub-dominios llamados manos de EF, los cuales representan sitios de enlace para calcio. El gen de la centrina humana 2 está codificado en el cromosoma X y ha sido nombrado como Hcetn2. Hcen 2 parece desempeñar un papel importante en la duplicación y segregación del centrosoma durante la mitosis. Células bacterianas competentes E. coli fueron transformadas con el gen que codifica para Hcetn2 y crecidas en dos tipos de caldos de cultivo: 2xYT y en un medio mínimo enriquecido marcado con 13C. Luego se indujeron para sobre-expresar las proteínas Hcen 2 y 13C-Hcen 2. Dichas proteínas fueron purificadas usando cromatografía de afinidad hidrofóbica, la cual fue seguida por varios pasos de separación mediante cromatografía de intercambio aniónico. La centrina humana 2 y su forma marcada con 13C fueron obtenidas con gran pureza para realizar estudios biofísicos. Los estudios de calorimetría de rastreo diferencial de la centrina humana 2 muestran que esta es una proteína estable con una pre-transición a 85 °C y una temperatura de desnaturalización de 120 °C.en_US
dc.description.sponsorshipMayo clinic and Foundation DNA core facility, the Protein core facility and to Vanderbilt University Mass Spectral Facilityen_US
dc.language.isoenen_US
dc.subjectHuman centrin 2 (Hcen 2)en_US
dc.subjectEF-handsen_US
dc.subjectHcetn2en_US
dc.subjectBacterial E. coli cellsen_US
dc.subjectHydrophobic affinity chromatographyen_US
dc.subjectC-homogenously labeled proteinen_US
dc.subject.lcshCalcium-binding proteins.en_US
dc.subject.lcshCentrioles.en_US
dc.titleExpression, purification and characterization of a human calcium-binding protein: Human centrin 2en_US
dc.typeThesisen_US
dc.rights.licenseAll rights reserveden_US
dc.rights.holder(c) 2005 Verónica Meza Venenciaen_US
dc.contributor.committeeAlfaro, Mónica
dc.contributor.committeeDiffoot-Carlo, Nanette
dc.contributor.committeeRios Steiner, Jorge
dc.contributor.representativeVillafañe, Gilberto
thesis.degree.levelM.S.en_US
thesis.degree.disciplineBiologyen_US
dc.contributor.collegeCollege of Arts and Sciences - Sciencesen_US
dc.contributor.departmentDepartment of Biologyen_US
dc.description.graduationYear2005en_US


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