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HPLC elution profiles of purified mutant R3s. Purified I155L (a), I155L/I181L (b), and I155L/I181V (c) were applied to a Cosmosil 5C4-300 column (Nacalai Tesque, Kyoto) and equilibrated with 0.1% trifluoroacetic acid. The proteins were eluted with a linear gradient of acetonitrile (0–60%) in 0.1% trifluoroacetic acid.
