Expression and Purification of Inflammasome Sensor NOD-Like Receptor Protein-1 Using the Baculovirus-Insect Cell
Hariharan Sivaraman1,2, Wilson Wong3,4
1Centre for Innate Immunity and Infectious Diseases, Centre for Cancer Research, Hudson Institute of Medical Research, Melbourne, Clayton, VIC, Australia.
Abstract:
Inflammasomes are innate immune sensing and signaling complexes critical for defense against pathogens and response to cellular stresses. A core component of inflammasomes is the sensor protein, which, upon sensing pathogen- or danger-associated molecular patterns (PAMPs or DAMPs), converts from inactive to active signaling platform for initiation of inflammatory signaling. A reliable source for the production and purification of recombinant inflammasome sensors is therefore invaluable for biochemical and structural characterizations, as well as drug screening for the development of therapeutics. Here, we describe an expression and purification protocol using the baculovirus-insect cell expression system to generate recombinant NLRP1, an important member of the NOD-like receptor (NLR) family of inflammasome sensors.
