RESUMEN
The metabolic utilization of substrates results in ATP synthesis and energy loss as heat. In tissues and cells the mitochondria reoxidize reduced coenzymes and phosphorylate ADP. A significant proportion of the energy is released through thermogenesis by mitochondria. This is due to a less than perfect coupling of cellular respiration to ATP synthesis. Previous studies of brown adipocytes, which are cells specialized in regulatory thermogenesis, have shown that heat production is due to the regulated activity and synthesis of a particular proton transporter in the inner membrane of brown adipocyte mitochondria--uncoupling protein (UCP) 1. UCP homologues have recently been identified. UCP2 is widely expressed in human tissues, whereas UCP3 is expressed predominantly in human skeletal muscles. These novel UCPs represent genes which are potentially important for regulation of metabolic pathways and energy expenditure in humans. Biochemical and genetic studies support a role for these novel UCPs in metabolic regulations in humans. However, several physiological studies question such a role. Importantly, UCP2 and UCP3 seem to be able to control the activity of mitochondria in response to oxidants.
Asunto(s)
Proteínas Portadoras/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas de Transporte de Membrana , Mitocondrias/metabolismo , Proteínas Mitocondriales , Proteínas/metabolismo , Termogénesis/fisiología , Metabolismo Energético/fisiología , Homeostasis/fisiología , Humanos , Canales Iónicos , Investigadores , Proteína Desacopladora 1 , Proteína Desacopladora 2 , Proteína Desacopladora 3Asunto(s)
Tejido Adiposo Pardo/metabolismo , Proteínas Portadoras/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas de Transporte de Membrana , Proteínas Mitocondriales , Desacopladores/metabolismo , Secuencia de Aminoácidos , Animales , Metabolismo Basal/genética , Proteínas Portadoras/química , Proteínas Portadoras/genética , Clonación Molecular , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Canales Iónicos , Proteínas de la Membrana/química , Proteínas de la Membrana/genética , Mitocondrias/metabolismo , Proteínas Desacopladoras Mitocondriales , Modelos Moleculares , Plantas , Proteínas/química , Proteínas/metabolismo , Retinoides/farmacología , Homología de Secuencia , Desacopladores/química , Proteína Desacopladora 1 , Proteína Desacopladora 2 , Proteína Desacopladora 3 , Levaduras/genética , Levaduras/metabolismoRESUMEN
The cDNA of an uncoupling protein (UCP) homologue was obtained by screening a chicken skeletal-muscle library. The predicted 307-amino-acid sequence of avian UCP (avUCP) is 55, 70, 70 and 46% identical with mammalian UCP1, UCP2 and UCP3 and plant UCP respectively. avUCP mRNA expression is restricted to skeletal muscle and its abundance was increased 1.3-fold in a chicken line showing diet-induced thermogenesis, and 3.6- and 2.6-fold in cold-acclimated and glucagon-treated ducklings developing muscle non-shivering thermogenesis respectively. The present data support the implication of avUCP in avian energy expenditure.
Asunto(s)
Proteínas Aviares , Proteínas Portadoras/fisiología , Proteínas Mitocondriales , Músculo Esquelético/fisiología , Termogénesis/fisiología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Northern Blotting , Proteínas Portadoras/química , Proteínas Portadoras/genética , Pollos , Cartilla de ADN , ADN Complementario , Proteínas Desacopladoras Mitocondriales , Datos de Secuencia Molecular , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Homología de Secuencia de AminoácidoRESUMEN
Uncoupling protein 2 (UCP2) belongs to the mitochondrial anion carrier family and partially uncouples respiration from ATP synthesis when expressed in recombinant yeast mitochondria. We generated a highly sensitive polyclonal antibody against human UCP2. Its reactivity toward mitochondrial proteins was compared between wild type and ucp2(-/-) mice, leading to non-ambiguous identification of UCP2. We detected UCP2 in spleen, lung, stomach, and white adipose tissue. No UCP2 was detected in heart, skeletal muscle, liver, and brown adipose tissue. The level of UCP2 in spleen mitochondria is less than 1% of the level of UCP1 in brown adipose tissue mitochondria. Starvation and LPS treatments increase UCP2 level up to 12 times in lung and stomach, which supports the hypothesis that UCP2 responds to oxidative stress situations. Stimulation of the UCP2 expression occurs without any change in UCP2 mRNA levels. This is explained by translational regulation of the UCP2 mRNA. We have shown that an upstream open reading frame located in exon two of the ucp2 gene strongly inhibits the expression of the protein. This further level of regulation of the ucp2 gene provides a mechanism by which expression can be strongly and rapidly induced under stress conditions.
Asunto(s)
Proteínas de Transporte de Membrana , Proteínas Mitocondriales , Estrés Oxidativo , Biosíntesis de Proteínas , Proteínas/metabolismo , Animales , Secuencia de Bases , Células COS , Cartilla de ADN , Exones , Humanos , Canales Iónicos , Ratones , Ratones Noqueados , Sistemas de Lectura Abierta , Proteínas/genética , ARN Mensajero/genética , Ratas , Proteína Desacopladora 2RESUMEN
The gene Ucp2 is a member of a family of genes found in animals and plants, encoding a protein homologous to the brown fat uncoupling protein Ucp1 (refs 1-3). As Ucp2 is widely expressed in mammalian tissues, uncouples respiration and resides within a region of genetic linkage to obesity, a role in energy dissipation has been proposed. We demonstrate here, however, that mice lacking Ucp2 following targeted gene disruption are not obese and have a normal response to cold exposure or high-fat diet. Expression of Ucp2 is robust in spleen, lung and isolated macrophages, suggesting a role for Ucp2 in immunity or inflammatory responsiveness. We investigated the response to infection with Toxoplasma gondii in Ucp2-/- mice, and found that they are completely resistant to infection, in contrast with the lethality observed in wild-type littermates. Parasitic cysts and inflammation sites in brain were significantly reduced in Ucp2-/- mice (63% decrease, P<0.04). Macrophages from Ucp2-/- mice generated more reactive oxygen species than wild-type mice (80% increase, P<0.001) in response to T. gondii, and had a fivefold greater toxoplasmacidal activity in vitro compared with wild-type mice (P<0.001 ), which was absent in the presence of a quencher of reactive oxygen species (ROS). Our results indicate a role for Ucp2 in the limitation of ROS and macrophage-mediated immunity.