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Compared to LPS, MPLA induced a qualitatively similar, but less potent pro-inflammatory immune response, but was unable to activate human or mouse MCs.

A novel lectin (SML-2) consisting of 138 amino acids was isolated from cyst merozoites of Sarcocystis muris and sequenced by Edman degradation and mass spectrometry. All 12 cysteinyl residues are involved in disul®de bridges, four of which are attributed to a characteristic pattern of cysteines as found in the so-called PANmodule superfamily. Crystals of SML-2 diffracting to 2.1 A Ê resolution at a synchrotron were grown by the hanging-drop vapour-diffusion technique. They belong to the space group P2 1 2 1 2 1 , with unit-cell parameters a = 53.6, b = 128.8, c = 158.2 A Ê and eight molecules in the asymmetric unit. SML-2 cocrystallized with Au galactose results in two different crystal forms. The ®rst form is isomorphous with the native crystals and the second form adopts space group C222 1 , with unit-cell parameters a = 74.7, b = 82.0, c = 131.0 A Ê , and diffracts to 2.4 A Ê at a rotating-anode X-ray generator.

The mature major microneme protein of Sarcocystis muris cyst merozoites, which is known as a dimeric lectin with high affinity to galactose and some of its derivatives, was expressed in Escherichia colias a histidine-tagged fusion protein. The recombinant polypeptide, which was recognized by a monoclonal antibody directed against the native lectin, was purified from inclusion bodies after solubilization and refolding, using a combination of metal chelate and lactose affinity chromatography. The apparent molecular mass of the refolded polypeptide as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoreses was 16 kDa, whereas gel filtration chromatography clearly demonstrated that the recombinant protein, like its native counterpart, exists as a homodimer of two non-covalently associated subunits. Inhibition of haemagglutination suggests that the combining site of the recombinant lectin recognizes N-acetyl-galactosamine as the dominant sugar, thus confirming the correct folding of the monosaccharide combining site in the renatured lectin. To the best of our knowledge, this work represents the first reported detailed characterization of a recombinant lectin from apicomplexan parasites, and may contribute to a better understanding of the process of host cell recognition and invasion by these obligate intracellular protozoa.