Apolipoprotein E4 heterologous expression, purification under non-denaturing conditions, and effects on neuronal clonal cell lines

dc.creatorSerrano, Ezequiel
dc.creatorBarrantes, Francisco José
dc.creatorValdivieso, Ángel Gabriel
dc.date2023-06-28T16:00:41Z
dc.date2023-06-28T16:00:41Z
dc.date2023
dc.date.accessioned2023-08-30T15:41:01Z
dc.date.available2023-08-30T15:41:01Z
dc.identifierSerrano, E., Barrantes, F.J., Valdivieso, A.G. Apolipoprotein E4 heterologous expression, purification under non-denaturing conditions, and effects on neuronal clonal cell lines [en línea]. Protein Expression and Purification. 2023, 210, doi:10.1016/j.pep.2023.106312 Disponible en: https://repositorio.uca.edu.ar/handle/123456789/16628
dc.identifier1046-5928
dc.identifierhttps://repositorio.uca.edu.ar/handle/123456789/16628
dc.identifier10.1016/j.pep.2023.106312
dc.identifier37236517
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8526770
dc.descriptionAbstract: The ε4 allele of the apolipoprotein E gene (APOE4) constitutes the main genetic risk factor for late-onset Alzheimer disease (AD). High amounts of pure apolipoprotein E4 (ApoE4), in a rapid and reproducible fashion, could be of value for studying its pathophysiological roles in AD. The aim of the present work was to optimize a preparative method to obtain highly purified recombinant ApoE4 (rApoE4) with full biological activity. rApoE4 was expressed in the E. Coli BL21(D3) strain and a soluble form of the protein was purified by a combination of affinity and size-exclusion chromatography that precluded a denaturation step. The structural integrity and the biochemical activity of the purified rApoE4 were confirmed by circular dichroism and a lipid-binding assay. Several biological parameters affected by rApoE4, such as mitochondrial morphology, mitochondrial membrane potential and reactive oxygen species production were studied in CNh cells, a neuronal cell line, and neurodifferentiation and dendritogenesis were analyzed in the SH-SY5Y neuroblastoma cell line. The improved rApoE4 purification technique reported here enables the production of highly purified protein that retain the structural properties and functional activity of the native protein, as confirmed by tests in two different neuronal cell lines in culture.
dc.formatapplication/pdf
dc.languagespa
dc.publisherElsevier
dc.rightsAcceso Restringido
dc.rightshttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.sourceProtein Expression and Purification. 2023, 210
dc.subjectENFERMEDAD DE ALZHEIMER
dc.subjectPROTEINAS
dc.subjectAPOLIPOPROTEINA
dc.titleApolipoprotein E4 heterologous expression, purification under non-denaturing conditions, and effects on neuronal clonal cell lines
dc.typeArtículo


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