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Browsing by Subject "BRET"

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    Open Access
    A genetically encoded IL-1β BRET sensor to monitor inflammasome activity
    (Asociación Americana de Inmunología, 2012) Compan, Vincent; Baroja-Mazo, Alberto; Bragg, Laricia; Verkhratsky, Alexei; Perroy, Julie; Pelegrin, Pablo; Bioquímica y Biología Molecular B e Inmunología
    Inflammation is fundamental for protecting the organism against infection and injury. However, a failure to control immune response results in chronic inflammation and several associated disorders such as pain and loss of function. Initiation of inflammation is orchestrated by cytokines, among which interleukin (IL)-1β is particularly important. IL- 1β is synthesized as an inactive protein that has to be processed by the inflammasome to generate the mature bioactive form. Conventional techniques cannot monitor IL-1β activation with high spatial and temporal resolution. Here, we present a ratiometric biosensor that allows monitoring IL-1β processing in real-time, with a temporal resolution of seconds and with a single cell spatial resolution. Using this sensor, we describe for the first time the kinetic of the inflammasome activity in living macrophages. With this new probe we also demonstrated that the pro-IL-1β processing occurs all over the cytoplasm.

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