Photooxidation of dinitrophenylhistidine-200 human carbonic anhydrase B.

Partial inactivation of tau-dinitrophenylhistidine-200 human carbonic anhydrase B, induced by visible light, followed first order kinetics (k(app) = 6.05 times 10-2 min-1). After 50 min the tau-dinitrophenylhistidine (tau-DNP-histidine) content decreased to a negligible level, but the illuminated enzyme retained, at pH 7.6, approximately 9.2 percent of the esterase activity of the native enzyme. The following lines of evidence suggest that the loss of activity results from the destruction of tau-DNP-histidine-200. (1) No significant loss of amino acid other than tau-DNP-histidine was detected after illumination. (2) The rate of loss of activity correlated well with the loss of tau-DNP-histidine. (3) In the photooxidized enzyme the DNP moiety was retained but had lost the characteristic sensitivity of tau-DNP-histidine to nucleophilic attack. Titration of the illuminated enzyme with acetazolamide indicated that the residual activity is an intrinsic property of the modified enzyme. The chromatographically purified photooxidized enzyme migrated as a single band on isoelectrofocusing in polyacylamide gel, and at pH 7.6 possessed 7.5 percent esterase activity relative to the native enzyme. By establishing effective destruction of histidine-200, it can be concluded that neither the pi N nor, as previously shown, the tau N of histidine-200 is critical for the catalysis.     

DDT induced ethological changes in estuarine fish

Synopsis Experiments were carried out to study the feeding rates of the predator fish Therapon jarbua (Forsk) on mullet juveniles, before and after treatment with DDT. Mullet juveniles treated with a subacute concentration, were refused by the control predators, whereas predators treated with a subacute concentration consumed more mullet juveniles. In the present study crescent perch T. jarbua were exposed to subacute and acute concentrations of DDT, and their behaviour was compared with that of the control predators. There were changes in oriented behaviour and co-ordinated movements, and in feeding, aggression and comfort behaviour of the fish. Inflammation in the gills, and caudal fin serration, were noticed in treated fishes. The findings presented here throw light on fundamental pathways by which pollutants interact with the behaviour of fishes.     

Activity of the respiratory inhibitor RF in human red cells in anaemia

Stroma-free haemolysates of red cells from patients with acute or latent anaemia inhibit the NADH oxidase activity of submitochondrial particles from beef heart. The inhibitory activities appeared significantly more frequently in anaemic patients than in normal persons and are likely due to the action of a factor which is identical or similar to the inhibitory protein RF present in rabbit reticulocytes. The possible use of the inhibitor assay for clinical purpose is under discussion.     

Mouse prolactin obtained by pituitary organ culture in a serum-free medium.

Mouse prolactin was purified by organ culture of pituitaries and electrophoresis of the medium. Mouse pituitaries were organ-cultured in 9-cm Petri dishes containing Waymouth's medium (MB 752/1) supplemented with penicillin (50 units/ml), streptomycin (100mug/ml), and bovine insulin (0.12 I.U./ml) for 8 days. Prolactin-rich culture medium was half-saturated with ammonium sulfate and centrifuged. The pellet was subjected to analytical disc electrophoresis (10% acrylamide). Gels were sectioned into 2-mm segments. Prolactin was eluted in 0.04M phosphate buffer (pH 7.2), dialyzed and lyophylized. Two hundred and forty ml medium in which 360 pituitaries were cultured yielded 29.3 mg lyophylized mouse prolactin. Although the preparation contained 2 other bands on acrylamide gel disc electrophoresis, a single precipitin line was seen in immunodiffusion and immunoelectrophoresis, showing the identity of their antigenicity. From these results, two other proteins in the preparation were suggested to be deamidated prolactin.