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. 2015 Jul 14;4(5):743–752. doi: 10.1002/mbo3.275

Figure 2.

Figure 2

Kinetics of SprP activation. (A) Purified SprP-HylA1 fusion protein was incubated at 8°C and enzymatic activity tested with resorufin-labeled casein as the substrate. (B) SprP in 200 mmol/L Tris-HCl, 5 mmol/L CaCl2, pH 8 was 10-fold concentrated by TCA precipitation and a 10 μL aliquot was analyzed by SDS-PAGE; S = Mr standard proteins (PageRuler Plus Prestained Protein Ladder, Fermentas, Germany). (C) One microgram of enzymatically active SprP was subjected to casein gel zymography. White arrows indicate proteins showing protease activity corresponding to Mr of 88 kDa (SprP-HlyA1 fusion protein) and 26 kDa (mature SprP). S = Mr standard proteins (Precision Plus Protein Dual Color Standard, Bio-Rad, Germany). TCA, trichloroacetic acid; SDS-PAGE, sodiumdodecyl sulfate polyacrylamide gel electrophoresis.