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dc.contributor.advisorPastrana-Ríos, Belinda
dc.contributor.authorDe Orbeta-Cruz, Jessica M.
dc.date.accessioned2018-05-16T16:34:48Z
dc.date.available2018-05-16T16:34:48Z
dc.date.issued2006
dc.identifier.urihttps://hdl.handle.net/20.500.11801/594
dc.description.abstractHuman centrin 1 (Hcen 1) is an acidic calcium binding protein with a molecular mass of ~ 20 kDa and has been localized in human as well as in mouse testis, at the base of the flagellar apparatus in sperm cells. Its putative function is associated with the fertilized zygote duplication and motility of the sperm. In an effort to provide a better understanding of the Hcen 1 structure, high level expression and protein purification, using affinity and anion exchange chromatography, were performed. A successful expression was obtained with optical density measures of 9 and an average of 40 grams of bacterial pellet, what leads to 10 mg of ~ 99% pure Hcen 1. Differential Scanning Calorimetry studies were done using the purified Hcen 1, obtaining two pre-transitional peaks at ~ 58.42 ûC and ~ 88.82 ûC. These results suggest that Hcen 1 undergoes conformational changes related with its N and C- terminal calcium binding capacities.en_US
dc.description.abstractLa isoforma humana centrin 1 (Hcen 1) es una proteína acídica con capacidad para enlazar moléculas de calcio. Esta proteína tiene un peso molecular de ~ 20 kDa y ha sido localizada en el cuerpo basal del flagelo del espermatozoide. Se ha propuesto que su función esta asociada a la primera división mitótica del cigoto y al motilidad del espermatozoide. En un esfuerzo por proveer un mejor entendimiento sobre la estructura de Hcen 1 se realizaron experimentos de expresión a gran escala seguidos de dos pasos de purificación utilizando cromatografía de afinidad e intercambio iónico. Se obtuvo una densidad óptica máxima de 9 y un promedio de 40 gramos de células de bacteria, finalmente se obtuvieron 10 mg de Hcen 1 con una pureza de ~ 99%. Estudios empleando la técnica de Calorimetría Diferencial de Barrido fueron realizados utilizando muestra de la proteína purificada, dos pre transiciones fueron observadas en Hcen 1 a ~ 58.42 ûC y ~ 88.82 ûC, esto resultados sugieren cambios conformacionales en la proteína que podrían ser explicados por la diferencia existente en la capacidad de enlace de moléculas de calcio entre el terminal amino y carboxilo.en_US
dc.description.sponsorshipNIH MBRS-SCORE S06GM08103en_US
dc.language.isoenen_US
dc.subjectHuman centrin 1 (Hcen 1)en_US
dc.subjectAcidic calcium binding proteinen_US
dc.subject.lcshCalcium-binding protein genes--Expression.en_US
dc.titleHigh level expression and thermal characterization of human centrinen_US
dc.typeThesisen_US
dc.rights.licenseAll rights reserveden_US
dc.rights.holder(c) 2006 Jessica M. De Orbeta-Cruzen_US
dc.contributor.committeeDiffoot - Carlo, Nanette
dc.contributor.committeeMontalvo - Rodríguez, Rafael
dc.contributor.representativeRíos, Jorge
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.graduationYear2006en_US


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