Meza-Venencia, Verónica

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Expression, purification and characterization of a human calcium-binding protein: Human centrin 2
(2005) Meza-Venencia, Verónica; Pastrana-Ríos, Belinda; College of Arts and Sciences - Sciences; Alfaro, Mónica; Diffoot-Carlo, Nanette; Rios Steiner, Jorge; Department of Biology; Villafañe, Gilberto
Human 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.