Asunto(s)
Apoptosis , Ciclooxigenasa 2/metabolismo , Fibroblastos/enzimología , Proteínas Represoras/metabolismo , Animales , Caspasas/metabolismo , Células Cultivadas , Ciclooxigenasa 2/deficiencia , Ciclooxigenasa 2/genética , Electroforesis en Gel Bidimensional , Embrión de Mamíferos , Fibroblastos/patología , Regulación del Desarrollo de la Expresión Génica , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Ratones , Ratones Noqueados , Prohibitinas , ARN Mensajero/metabolismo , Proteínas Represoras/genética , Reproducibilidad de los Resultados , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Factores de Tiempo , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
Atopic dermatitis (AD) is a chronic inflammatory skin disease that induces changes in various inflammatory skin cells. The prevalence of AD is as high as 18% in some regions of the world, and is steadily rising. However, the pathophysiology of AD is poorly understood. To identify the proteins involved in AD pathogenesis, a comparative proteomic analysis of protein expression in peripheral blood mononuclear cells isolated from AD patients and healthy donors was conducted. Significant changes were observed in the expressions of fourteen proteins, including the vinculin, PITPNB, and Filamin A proteins. Among the proteins, alpha-SNAP and FLNA decreased significantly, and PITPNB increased significantly in AD patients compared with control subjects; these findings were further confirmed by real-time PCR and Western blot analysis. The comparative proteome data may provide a valuable clue to further understand AD pathogenesis, and several differentially regulated proteins may be used as biomarkers for diagnosis and as target proteins for the development of novel drugs.
Asunto(s)
Dermatitis Atópica/sangre , Leucocitos Mononucleares/metabolismo , Secuencia de Bases , Biomarcadores/sangre , Proteínas Sanguíneas/genética , Proteínas Sanguíneas/aislamiento & purificación , Proteínas Sanguíneas/metabolismo , Estudios de Casos y Controles , Cartilla de ADN/genética , Dermatitis Atópica/etiología , Dermatitis Atópica/genética , Electroforesis en Gel Bidimensional , ProteómicaRESUMEN
Atopic dermatitis (AD) is an inflammatory skin disorder that is both uncomfortable and distressing to patients, and its prevalence has been steadily increasing. It is obvious that the identification of efficient markers of AD in plasma would offer the possibility of effective diagnosis, prevention, and treatment strategies. In this study, a proteomic approach was used to analyze plasma glycoproteins from both children with AD and healthy child donors. Several protein spots showing significant quantitative changes in the AD patients were identified. Through sequential studies, it was confirmed that CD5L and ApoE were significantly up-regulated or down-regulated, respectively, in the plasma from AD patients compared with that from healthy donors. In addition, we suggest that the up-regulated CD5L in AD patients causes eosinophilia by inhibiting apoptosis or promoting the proliferation of eosinophils either in combination with or without IL-5. The glycoproteomic data in this study provides clues to understanding the mechanism of atopic alterations in plasma and suggests AD-related proteins can be used as candidate markers for AD.
Asunto(s)
Apolipoproteínas E/sangre , Dermatitis Atópica/metabolismo , Glicoproteínas/sangre , Receptores Depuradores de Clase B/sangre , Proteínas Reguladoras de la Apoptosis , Biomarcadores/sangre , Línea Celular , Proliferación Celular , Niño , Eosinofilia/metabolismo , Eosinófilos/fisiología , Femenino , Humanos , Interleucina-5/metabolismo , Masculino , Proteómica , Receptores DepuradoresRESUMEN
Oxidative stress is one of the major causes of neuronal cell death in disorders such as perinatal hypoxia and ischemia. Protein phosphorylation is the most significant PTM of proteins and plays an important role in stress-induced signal transduction. Thus, the analysis of alternative protein phosphorylation states which occur during oxidative stress-induced cell death could provide valuable information regarding cell death. In this study, a reference phosphoproteome map of the mouse hippocampal cell line HT22 was constructed based on 125 spots that were identified by MALDI-TOF or LC-ESI-Q-TOF-MS analysis. In addition, proteins of HT22 cells at various stages of oxidative stress-induced cell death were separated by 2-DE and alterations in phosphoproteins were detected by Pro-Q Diamond staining. A total of 17 spots showing significant quantitative changes and seven newly appearing spots were identified after glutamate treatment. Splicing factor 2, peroxiredoxin 2, S100 calcium binding protein A11, and purine nucleoside phosphorylase were identified as up- or down-regulated proteins. CDC25A, caspase-8, and cyp51 protein appeared during oxidative stress-induced cell death. The data in this study from phosphoproteomic analysis provide a valuable resource for the understanding of HT22 cell death mechanisms mediated by oxidative stress.
Asunto(s)
Apoptosis/efectos de los fármacos , Ácido Glutámico/farmacología , Neuronas/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Proteoma/análisis , Animales , Muerte Celular/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Ácido Edético/farmacología , Ácido Egtácico/farmacología , Hipocampo/citología , Espectrometría de Masas , Ratones , Neuronas/metabolismo , Fosfoproteínas/análisis , Fosfoproteínas/metabolismo , Fosforilación , Proteómica/métodos , Factores de TiempoRESUMEN
The roles that accessory gene products play in activating the Helicobacter pylori urease apoprotein were examined. The activity of the urease apoprotein increased in the following order when it was expressed with the accessory genes: ureGAsunto(s)
Apoproteínas/metabolismo
, Proteínas Bacterianas/metabolismo
, Regulación Bacteriana de la Expresión Génica
, Helicobacter pylori/enzimología
, Ureasa/metabolismo
, Apoproteínas/genética
, Proteínas Bacterianas/genética
, Proteínas Portadoras/genética
, Proteínas Portadoras/metabolismo
, Activación Enzimática
, Immunoblotting
, Proteínas de Unión a Fosfato
, Plásmidos
, Ureasa/genética