RESUMEN
OBJECTIVE: The purpose of the article is to evaluate the changes in lipid metabolism in bovine mammary-gland epithelial MAC-T cells after PKM2 knockdown. RESULTS: MAC-T cells stably expressing low levels of PKM2 were established with lentivirus-mediated small hairpin RNA. Although the knockdown of PKM2 had no effect on MAC-T cell growth, the reduced expression of PKM2 attenuated the mRNA and protein expression of key enzymes involved in sterol synthesis through the SREBP pathway. CONCLUSIONS: The downregulation of PKM2 significantly influenced lipid synthesis in bovine mammary-gland epithelial MAC-T cells. These findings extend our understanding of the crosstalk between glycolysis and lipid metabolism in bovine mammary-gland epithelial cells.
Asunto(s)
Proteínas Portadoras/genética , Metabolismo de los Lípidos/genética , Glándulas Mamarias Animales/metabolismo , Proteínas de la Membrana/genética , Proteínas de Unión a los Elementos Reguladores de Esteroles/genética , Hormonas Tiroideas/genética , Animales , Proteínas Portadoras/metabolismo , Bovinos , Células Epiteliales/metabolismo , Femenino , Técnicas de Silenciamiento del Gen , Glucólisis/genética , Lípidos/biosíntesis , Proteínas de la Membrana/metabolismo , ARN Mensajero/genética , Transducción de Señal , Proteínas de Unión a los Elementos Reguladores de Esteroles/metabolismo , Linfocitos T/metabolismo , Hormonas Tiroideas/metabolismo , Proteínas de Unión a Hormona TiroideRESUMEN
Based on the complexation between proteins and Cu(II) coupled with the time-resolved chemiluminescence (CL) technique, a highly sensitive and quantitative assay for measuring proteins in solution is described. The complexes of proteins with Cu(II) have a strongly catalytic effect on the luminol-H2O2 CL reaction. Because the CL emission produced by the complexes is much more long-lived than that by Cu(II), the CL signals originating from proteins can be easily identified and measured with a time-resolved technique. On this basis, bovine albumin fraction V (BAF V) can be quantitatively determined in the range of 0.01 - 5.0 microg/ml with a detection limit of 5.8 ng/ml. The results show that the proposed assay exhibits a small variation in the response values for the same amount of different proteins, as compared to the Lowry as well as Bradford assays. The CL assay has also been studied for the detection of immobilized proteins.