Análisis funcional de mutantes genéticos de la tropomiosina humana
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2016
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Jaén: Universidad de Jaén
Resumen
[ES]Una nueva mutación en el gen de la tropomiosina humana 1 (TPM1) ha sido identificada en un paciente diagnosticado con Cardiomiopatía Hipertrófica en el Hospital Regional “Ciudad de Jaén”. En este trabajo planteamos que dicho cambio de ácido glutámico por lisina en la posición 62 de la secuencia peptídica (E62K) podría ser disfuncional y por tanto, podría ser causativa de dicha enfermedad. Dado que una mutación en la misma posición del gen TPM1, pero afectando de modo diferente (E62Q) ha sido previamente identificada en una familia con esta misma patología, hemos decidido estudiar sus efectos funcionales de forma paralela. Para ello, cimentamos nuestros estudios experimentales sobre la técnica de mutagénesis dirigida con la que hemos introducido de forma selectiva cambios nucleotídicos (E62K y E62Q respectivamente). La transfección de estos plásmidos en células cardiomiocíticas HL-1 nos ha permitido revelar que la mutación E62K, pero no la E62Q, provoca un aumento de volumen celular. Además, estas alteraciones se agravan en presencia de un agente pro-hipertrófico, como la angiotensina II. Por tanto, nuestros resultados muestran que la mutación E62K, pero no la E62Q, provoca cambios funcionales en la proteína TPM1.
[EN]A new mutation in the gene coding for human tropomyosin 1 (TPM1) has been identified in a patient diagnosed with Hypertrophic Cardiomyopathy at the Regional Hospital “Ciudad de Jaén”. In this study we proposed that the change of glutamic acid for lysine at position 62 of the peptide sequence (E62K) could be dysfunctional, therefore could be causative of the disease. Since a mutation in the same gene position but affecting differently (E62Q) has previously been identified in a family with the same cardiac condition, we decided to study in parallel its putative functional defects. For this purpose, we have structured our experimental studies using site-directed mutagenesis technique by which we have selectivey introduced the corresponding nucleotide changes (E62K and E62Q respectively). Transient transfection assays using these plasmids into cardiomyocells HL-1 allowed us to reveal the E62K mutation, but not E62Q, causes increased cell volumen, a response which is further enhanced after treatment with pro-hypertrophy agents such as angiotensin II. Thus, our results show that E62K-mutation, but not E62Q, causes functional changes in the TPM1 protein.
[EN]A new mutation in the gene coding for human tropomyosin 1 (TPM1) has been identified in a patient diagnosed with Hypertrophic Cardiomyopathy at the Regional Hospital “Ciudad de Jaén”. In this study we proposed that the change of glutamic acid for lysine at position 62 of the peptide sequence (E62K) could be dysfunctional, therefore could be causative of the disease. Since a mutation in the same gene position but affecting differently (E62Q) has previously been identified in a family with the same cardiac condition, we decided to study in parallel its putative functional defects. For this purpose, we have structured our experimental studies using site-directed mutagenesis technique by which we have selectivey introduced the corresponding nucleotide changes (E62K and E62Q respectively). Transient transfection assays using these plasmids into cardiomyocells HL-1 allowed us to reveal the E62K mutation, but not E62Q, causes increased cell volumen, a response which is further enhanced after treatment with pro-hypertrophy agents such as angiotensin II. Thus, our results show that E62K-mutation, but not E62Q, causes functional changes in the TPM1 protein.