Nature research paper: Direct activation of a bacterial innate immune system by a viral capsid protein
-SUMO-gp57 and grown in LB medium toof 0.6. Expression of His–SUMO–Gp57 was induced by addition of 0.1 mM IPTG, and cells were grown for overnight at 16 °C. The subsequent purification, SUMO-tag cleavage and purity assessment steps were identical to the workflow described above for the all the CapRelprotein variants. After removing the SUMO tag, samples were concentrated to 10 μM and used directly for ITC immediately after purification.
Smode . Leucine enkephalin was applied for mass accuracy correction and sodium iodide was used as calibration for the mass spectrometer. Sdata were collected by a 20–30 V transfer collision energy ramp. The pepsin column was washed between injections using pepsin wash buffer methanol, 0.8% formic acid). A blank run was performed between each sample to prevent significant peptide carry-over. Optimized peptide identification and peptide coverage for all samples was performed from undeuterated controls . All deuterium time points were performed in triplicate.
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