Leslie B. Poole

Hydroxyapatite chromatography: altering the phosphate-dependent elution profile of protein as a function of pH.

Hydroxyapatite is a form of calcium phosphate thathaslongbeenusedinthechromatographicseparationofproteinsandDNA[1].Hydroxyapatiteisbestknownasacrystallinematerialbutisnowavailableinarangeofce-ramicderivativesthatarevastlysuperiorintermsofflowrate, stability, and reproducibility over many cycles ofuse.Thesedevelopmentshaveledtoarenewedinterestintheuseofthismediawithuniqueseparationproperties.Thisreportaimstofurtherextendthe usefulnessofhy-droxyapatiteforthepurificationofproteins.The adsorption of proteins to hydroxyapatite iscomplicated because it involves both anionic and cat-ionicexchange.TheCa

PEROXIDASE ACTIVITY OF A TSA-LIKE ANTIOXIDANT PROTEIN FROM A PATHOGENIC AMOEBA

The 29 kDa surface protein of Entamoeba histolytica is an abundant antigenic protein expressed by pathogenic strains of this organism. The protein is a member of a widely-dispersed group of homologues which includes at least two cysteinyl peroxidases, Salmonella typhimurium alkyl hydroperoxidase C-22 protein (AhpC) and Saccharomyces cerevisiae thiol-specific antioxidant protein (TSA). Here, for the first time in a pathogenic eukaryote, we have demonstrated that the amoebic protein also possesses peroxidatic and antioxidant activities in the presence of reductants such as dithiothreitol or thioredoxin reductase plus thioredoxin. Although the S. typhimurium AhpF flavoprotein was not an effective reductant of the amoebic TSA protein, one inhibitory monoclonal antibody directed toward amoebic TSA was also partially inhibitory toward reduced but not oxidized bacterial AhpC. These antioxidant proteins are likely to be important not only in general cell protection, but also in the promotion of infection and invasion by these pathogenic organisms through protection against oxidative attack by activated host phagocytic cells.