RESUMO
The use of the yeast Candida lipolytica as a support in affinity chromatography for the purification of lectins of Ricinus communis seeds has been studied. A comparison was made with columns of erythrocyte stromas and Sepharose 4B with respect to protein yields, the agglutinating activity yield, and the electrophoretic characteristics of the eluted substances. Even though the yield is less than with other supports, it does appear that the yeast columns represent an effective method for the purification of ricin lectins.
Assuntos
Candida , Cromatografia de Afinidade/métodos , Eletroforese em Gel de Poliacrilamida , Imunofluorescência , Testes de Hemaglutinação , Plantas Tóxicas , Ricinus , SefaroseRESUMO
Lectins in the seeds of Ricinus communis L. were separated by affinity chromatography using stromata, and 3 fractions obtained by ion-exchange chromatography. Polyacrylamide-gel electrophoresis showed that 2 fractions were homogeneous and that 1 fraction gave 2 bands. Immunoelectrophoresis with rabbit antisera and polyacrylamide-gel isoelectric focusing showed that the 3 fractions were separate and distinct entities consisting of many proteins with isoelectric points near the isoelectric point of the main protein. The molecular weights of the separated proteins and their subunits were studied by SDS polyacrylamide-gel electrophoresis.
Assuntos
Lectinas/isolamento & purificação , Proteínas de Plantas , Plantas Tóxicas , Ricinus , Animais , Cromatografia de Afinidade , Cromatografia por Troca Iônica , Testes de Hemaglutinação , Focalização Isoelétrica , Peso Molecular , Peptídeos , Lectinas de Plantas , CoelhosRESUMO
Stromata prepared with human or animal red blood cells are suspended in acrylamide solution. Gel as prepared for electrophoresis is dispersed, before use, and can be used in chromatographic columns for the retention of agglutinins. Lectins, e.g., where first absorbed and then eluted with solution of inhibitory sugar or with acid buffer. Some applications are mentioned.