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Separation and isolation of the hyaluronidase and phospholipase components of bee venom and investigation of bee venom-human serum interactions
Abstracts
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Bwxx~, S. A., MrrcHeaa-, A. W., WwL~rox, K. W. and Wr~rox, P. D., (Department of Chemistry, University of Birmingham, England) . Separation and isolation of the hyaluroni dase and phospholipase wmponents of bee venom and investigation of bee venom-human serum interactions . Clinics Chim. Acts 13, 582, 1966 . T~ wurxoxs purified the enzymes phospholipase A and hyaluronidase from bee venom and used them for m.-investigating bee venom-serum reactions. Purification was achieved by initial chromatography on a column of carboxymethyl Sephadex . This acidic ion exchange rosin binds the enzymes more loosely andRsz, Biochem.Z. 341, 451,196 . Forseparation of hyaluronithan the basic peptides (see H dase from phospholipase A, chromatography on alumina was used. Phospholipase A was homogeneous on cellulose acetate electrophoresis ; the purity of hyaluronidase was apparently not controlled . When whole human serum from normal subjects or beekeepers was allowed to diffuse against bee venom, precipitation lines appeared . The resemblance to true antigen antibody-precipitates was, however, superficial . Human serum was subjected first to agar gel electrophoresis, then to diffusion against bee venom then polyvalent antisenim then specific antisera against lipoproteins, so that it could be proved that bee venom reacts with lipoproteins, not with antibodies of senior . Purified hyaluronidase gave no rise to precipitation lines by gel diffusion, whereas phospholipase A was precipitating . For neutralization experiments, 7S~-globulins were produced by DEAE cellulose chromatography of normal and bcekeepers sera respectively . Hyaluronidase activity (viscosimetric test) was significantly diminished by preincubation with y-globulins from beekeepers but not from nonvals, whereas phospholipase A (degradation of lipoproteins in agar plates) was not inhibited. In sammarizing it has been demonstrated that man can produce neutralizing antibodies against bee venom hyaluronidase under circumstances of "physiological" envenomation, whereas the synthesis of antibodies against phospholipase A, clearly possible in rabbits (HAHBRMANN and Er. ICwxenn, Nature, Lond. 178, 1349, 1956) still awaits proof in man. E.H .
22 3
Bwxx~, S. A., MrrcHeaa-, A. W., WwL~rox, K. W. and Wr~rox, P. D., (Department of Chemistry, University of Birmingham, England) . Separation and isolation of the hyaluroni dase and phospholipase wmponents of bee venom and investigation of bee venom-human serum interactions . Clinics Chim. Acts 13, 582, 1966 . T~ wurxoxs purified the enzymes phospholipase A and hyaluronidase from bee venom and used them for m.-investigating bee venom-serum reactions. Purification was achieved by initial chromatography on a column of carboxymethyl Sephadex . This acidic ion exchange rosin binds the enzymes more loosely andRsz, Biochem.Z. 341, 451,196 . Forseparation of hyaluronithan the basic peptides (see H dase from phospholipase A, chromatography on alumina was used. Phospholipase A was homogeneous on cellulose acetate electrophoresis ; the purity of hyaluronidase was apparently not controlled . When whole human serum from normal subjects or beekeepers was allowed to diffuse against bee venom, precipitation lines appeared . The resemblance to true antigen antibody-precipitates was, however, superficial . Human serum was subjected first to agar gel electrophoresis, then to diffusion against bee venom then polyvalent antisenim then specific antisera against lipoproteins, so that it could be proved that bee venom reacts with lipoproteins, not with antibodies of senior . Purified hyaluronidase gave no rise to precipitation lines by gel diffusion, whereas phospholipase A was precipitating . For neutralization experiments, 7S~-globulins were produced by DEAE cellulose chromatography of normal and bcekeepers sera respectively . Hyaluronidase activity (viscosimetric test) was significantly diminished by preincubation with y-globulins from beekeepers but not from nonvals, whereas phospholipase A (degradation of lipoproteins in agar plates) was not inhibited. In sammarizing it has been demonstrated that man can produce neutralizing antibodies against bee venom hyaluronidase under circumstances of "physiological" envenomation, whereas the synthesis of antibodies against phospholipase A, clearly possible in rabbits (HAHBRMANN and Er. ICwxenn, Nature, Lond. 178, 1349, 1956) still awaits proof in man. E.H .