The I and S macromolecular fractions were suspended in 50 mM phosphate buffer at pH 7.0 to a final concentration of 20 mg/mL and treated with 40 UI/mL of Pronase E from Streptomyces griseus (4 UI/mg, Sigma-Aldrich, St. Louis, MI, USA) at 37°C for 24 h with magnetic stirring, and then centrifuged at 4,500 rpm for 30 min at 4°C. The supernatants obtained from the treatment of the I and S fractions were separated by centrifugation, freeze-dried, and labeled IHS and SH, respectively (Figure 1 ) (14 (link)).
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