Utilize este identificador para referenciar este registo: http://hdl.handle.net/10400.18/2467
Título: Advantages and Versatility of Fluorescence-Based Methodology to Characterize the Functionality of LDLR and Class Mutation Assignment
Autor: Etxebarria, A.
Benito-Vicente, A.
Alves, A.C.
Ostolaza, H.
Bourbon, M.
Martin, C.
Palavras-chave: Doenças Cardio e Cérebro-vasculares
Familial Hypercholesterolemia
Saúde Pública
Portugal
Data: 11-Nov-2014
Editora: Public Library of Science
Citação: PLoS One. 2014 Nov 11;9(11):e112677. doi: 10.1371/journal.pone.0112677. eCollection 2014.
Resumo: Familial hypercholesterolemia (FH) is a common autosomal codominant disease with a frequency of 1∶500 individuals in its heterozygous form. The genetic basis of FH is most commonly mutations within the LDLR gene. Assessing the pathogenicity of LDLR variants is particularly important to give a patient a definitive diagnosis of FH. Current studies of LDLR activity ex vivo are based on the analysis of 125I-labeled lipoproteins (reference method) or fluorescent-labelled LDL. The main purpose of this study was to compare the effectiveness of these two methods to assess LDLR functionality in order to validate a functional assay to analyse LDLR mutations. LDLR activity of different variants has been studied by flow cytometry using FITC-labelled LDL and compared with studies performed previously with 125I-labeled lipoproteins. Flow cytometry results are in full agreement with the data obtained by the 125I methodology. Additionally confocal microscopy allowed the assignment of different class mutation to the variants assayed. Use of fluorescence yielded similar results than 125I-labeled lipoproteins concerning LDLR activity determination, and also allows class mutation classification. The use of FITC-labelled LDL is easier in handling and disposal, cheaper than radioactivity and can be routinely performed by any group doing LDLR functional validations.
Peer review: yes
URI: http://hdl.handle.net/10400.18/2467
DOI: 10.1371/journal.pone.0112677
ISSN: 1932-6203
Aparece nas colecções:DPSPDNT - Artigos em revistas internacionais



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