RESUMO
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RESUMO
Aberrant expression of TNF family of cytokines has been linked to human diseases, and biologics targeting their signaling have become the best selling drugs globally. However, functional detection with labeled ligands for accurate detection of TNFR family of receptor-expressing target tissues or cell types remains to be developed. Here we show that TNF receptor family members are heat-stable and can be recognized both in vitro and in vivo by their ligands labeled with alkaline phosphatase. Such an approach may be used in lieu of antibodies for the identification of the cell types involved in receptor signaling during disease onset and progression.
Assuntos
Fosfatase Alcalina/metabolismo , Receptores do Fator de Necrose Tumoral/análise , Receptores do Fator de Necrose Tumoral/química , Marcadores de Afinidade , Animais , Células CHO , Cromatografia de Afinidade , Cricetulus , Células HCT116 , Humanos , Ligantes , Camundongos , Estabilidade Proteica , TermodinâmicaRESUMO
Background: Ascorbic acid (Asc) is one of the most abundant antioxidants and it serves as a major contributor to protect plants against oxidative damage. Plants use two enzymes that participate in the metabolic recycling of Asc. One of these two enzymes is dehydroascorbate reductase (DHAR). It directly regenerates Asc from its oxidized state and thus prevents Asc from being irreversibly hydrolyzed to 2, 3-diketogulonic acid. This study aimed to examine whether over-expression of DHAR leads to an enhanced oxidative stress tolerance in tobacco plants. Results: In this study, we functionally characterized a novel JcDHAR gene from Jatropha curcas and found via quantitative RT-PCR analysis that JcDHAR can be induced with H2O2, salt and PEG stresses. The DHAR activities of transgenic tobacco plants increased from 2.0 to 5.3 fold compared to wild-type plants. As a result, the transgenic plants displayed enhanced tolerance to oxidative stress. Conclusions: Our results indicate that JcDHAR expression can effectively enhance the tolerance to oxidative stress in plants.
