ABSTRACT
Apolipoprotein E (ApoE) is a major lipid carrier protein. In humans, ApoE is expressed in three polymorphic isoforms, which are encoded by three different alleles APOE2, APOE3, and APOE4. In the brains of Alzheimer's disease (AD) patients, each one of these three allelic isoforms is found in several "isoelectric" protein isoforms (qPI), i.e. protein isoforms resulting from PTMs altering the net charge (q) of the polypeptide. AD is a complex disease in which multiple causes and several risk factors affect the onset and disease outcome. A major risk factor for AD is ApoE4; therefore, it is important to characterize the different ApoE qPIs. We have implemented a detergent-based method for isolation and quantitation of protein isoforms, and we found differences in the solubility of protein isoforms depending on the type of solvent used. In this manuscript, we describe these methods and applied them to young human-ApoE targeted replacement mice. Our results indicate that there are no significant differences in the hippocampus proteome of these mice as a function of the APOE genotype.
Subject(s)
Apolipoprotein E3/biosynthesis , Apolipoprotein E4/biosynthesis , Proteome/analysis , Analysis of Variance , Animals , Apolipoprotein E3/analysis , Apolipoprotein E3/genetics , Apolipoprotein E3/metabolism , Apolipoprotein E4/analysis , Apolipoprotein E4/genetics , Apolipoprotein E4/metabolism , Creatine Kinase/analysis , Creatine Kinase/chemistry , Creatine Kinase/metabolism , Electrophoresis, Gel, Two-Dimensional , Genotype , Hippocampus/chemistry , Hippocampus/metabolism , Humans , Mice , Mice, Transgenic , Phosphopyruvate Hydratase/analysis , Phosphopyruvate Hydratase/chemistry , Phosphopyruvate Hydratase/metabolism , Protein Isoforms , Proteome/chemistry , Proteome/genetics , Proteome/metabolism , Proteomics/methods , SolubilityABSTRACT
En el presente artículo se presentan los conceptos básicos que definen la patología de la enfermedad de Alzheimer (EA) y los métodos fundamentales utilizados en Neuroproteómica para su estudio. De igual manera, se discuten algunos resultados en el análisis de esta enfermedad y su relación con el genotipo APOE4 de APOE, el gen que codifica la Apolipoproteína E (ApoE). Finalmente se hacen algunas consideraciones generales sobre la EA, cómo evitar su progresión y se discute brevemente el futuro de la investigación en esta área.
