The Toxoplasma adhesive protein MIC2 is proteolytically processed at multiple sites by two parasite-derived proteases

MIC2 is an adhesive protein that participates in host cell invasion by the obligate intracellular parasite Toxoplasma gondii, Earlier studies establis hed that MIC2 is secreted into the culture medium by extracellular parasite s and that release is coincident with proteolytic modification. Since littl e is known about proteolytic processing of proteins secreted by T. gondii, we undertook this study to investigate the proteolytic events that accompan y secretion of MICE. We demonstrate that the C-terminal domain of MICE is r emoved by a protease, termed MPP1, when MIC2 is released into the culture s upernatant. Additionally, prior to release, a second protease, termed MPP2, trims the N terminus of MIC2, resulting in the release of heterogeneously sized species of MIC2. Although MPP1 activity was unaffected by any of the protease inhibitors tested, MPP2 activity was blocked by a subset of serine and cysteine protease inhibitors. These results establish that MIC2 is pro teolytically modified at multiple sites by two distinct enzymes that probab ly operate on the parasite surface.