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 established that MIC2 is secreted into the culture medium by extracellular parasites and that release is coincident with proteolytic modification. Since little is known about proteolytic processing of proteins secreted by T. gondii, we undertook this study to investigate the proteolytic events that accompany secretion of MIC2. We demonstrate that the C-terminal domain of MIC2 is removed by a protease, termed MPP1, when MIC2 is released into the culture supernatant. 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 proteolytically modified at multiple sites by two distinct enzymes that probably operate on the parasite surface.
Carruthers
V B
VB
Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA. vcarruth@jhsph.edu
Sherman
G D
GD
Sibley
L D
LD
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D017931:DNA Primers; D006868:Hydrolysis; D016023:Integrins / Q000378:metabolism; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D014122:Toxoplasma / Q000378:metabolism*
D015853:Cysteine Proteinase Inhibitors; D017931:DNA Primers; D016023:Integrins; C408878:MIC2 protein, Toxoplasma gondii; D008565:Membrane Proteins; D015800:Protozoan Proteins; D015842:Serine Proteinase Inhibitors; D015638:Cytochalasin D; D012697:Serine Endopeptidases; D003546:Cysteine Endopeptidases