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. 2004 Aug;72(8):4836–4847. doi: 10.1128/IAI.72.8.4836-4847.2004

FIG. 3.

FIG. 3.

Proteolytic activation of proMMP-9 by SpeB as assessed by gelatinolytic activity. The gelatinolytic activity of MMP-9 produced after SpeB treatment was measured by using denatured type I collagen (gelatin) as a substrate for activated MMP-9. After 2 μM proMMP-9 was incubated with 2 μM SpeB, proMMP-9 treated with SpeB or untreated was incubated with gelatin (7 μg), followed by analysis for gelatin hydrolysis by use of SDS-10% PAGE. In some assays, E64 was added to the reaction mixture of proMMP with SpeB to inhibit SpeB proteinase activity. Lanes: (−), gelatin alone; proMMP-9, gelatin incubated with proMMP-9 alone; [proMMP-9+SpeB]→E64, gelatin incubated with SpeB-treated proMMP-9 after E64 treatment; and proMMP-9→[SpeB+E64], gelatin incubated with proMMP-9 treated with SpeB inactivated by E64. See the text for details.