title: A protease inhibitor associated with the surface of Toxoplasma gondii.
abstract: Toxoplasma gondii has a broad host-range including man and a variety of warm-blooded animals. The ability to infect and survive in this wide spectrum of hosts suggests highly evolved mechanisms to handle the harsh environments encountered. Here we show that extracellular tachyzoites are resistant to milligram levels of trypsin and describe the presence of an inhibitor of trypsin associated with the surface of T. gondii, TgTI. TgTI has an estimated molecular mass of 37000 dalton and is encoded by the TgTI-gene which is found at low abundance as an expressed sequence tag (EST) in both the bradyzoite and tachyzoite stages. The inhibitory binding region was found to be in the N-terminus of TgTI where aminoacid-alignment to earlier described protease inhibitors demonstrates 75% similarity. In functional analysis, recombinant TgTI-protein inhibits the activity of trypsin approximately 10 times more efficiently than an inhibitor isolated from soybean. In contrast to other known trypsin inhibitors, TgTI also possesses a predicted membrane-binding region. Polyclonal antibodies raised against recombinant TgTI bind to the surface of the tachyzoite stage as seen both by immunofluorescence and immunoprecipitation of surface labelled parasite proteins. The high survival rate of the parasite in the upper gastrointestinal tract may be enhanced by the presence of the TgTI-molecule.
authors.affiliation: Microbiology and Tumor Biology Center, Karolinska Institutet, Box 280, S-171 77, Stockholm, Sweden.
authors.forename: J G
authors.initials: JG
authors.lastname: Lindh
authors.forename: S
authors.initials: S
authors.lastname: Botero-Kleiven
authors.forename: J I
authors.initials: JI
authors.lastname: Arboleda
authors.forename: M
authors.initials: M
authors.lastname: Wahlgren
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D019084:Fluorescent Antibody Technique, Indirect; D008565:Membrane Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D015800:Protozoan Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000494:pharmacology; D016415:Sequence Alignment; D014122:Toxoplasma / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development; D014357:Trypsin; D014361:Trypsin Inhibitors / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology
chemical_list: D008565:Membrane Proteins; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D014361:Trypsin Inhibitors; D014357:Trypsin