Protein-carbohydrate-lipid complex isolated from the cell envelopes of Chlamydia psittaci in alkaline buffer and ethylenediaminetetraacetate.
Exposure of isolated cell envelopes from purified infectious elementary (EB) of Chlamydia psittaci to sodium carbonate-bicarbonate buffer at pH 10 plus ethylenediaminetetraacetate (EDTA) results in partial solubilization of the total protein. The released materials represent 20% of the dry weight, 16% of the total protein, 40% of the total carbohydrate, and 9% of the total lipid of the cell envelopes. Sucrose density gradient centrifugation, and Sephadex G-200, Sepharose 4B, or diethylaminoethyl-cellulose column chromatography, reveal a protein-carbohydrate-lipid complex of several hundred thousand molecular weight that contains 50% protein. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the isolated EB cell envelopes reveals two major protein bands, A and B, with estimated molecular masses of approximately 85,000 and 53,000, respectively, both of which also stain for the presence of carbohydrate and lipid. Gel electrophoresis of the protein-carbohydrate-lipid complex reveals two protein bands, C and D, with estimated molecular weights of approximately 17,000 and 13,000, respectively, which contain lipid and a small amount of carbohydrate; bands A and B are not present in the complex. Gel electrophoresis of the cell envelope residues after extraction of the complex with alkali and EDTA shows a single main band, corresponding to the position of band B, which contains protein, carbohydrate, and lipid; band A is completely missing. B and A is believed to be a component of the complex, which is split into two subunits on alkali solubilization.
D001426:Bacterial Proteins / Q000032:analysis; D002021:Buffers; D002241:Carbohydrates / Q000032:analysis; D002254:Carbonates; D002473:Cell Wall / Q000032:analysis / Q000187:drug effects / Q000648:ultrastructure; D002691:Chlamydophila psittaci / Q000648:ultrastructure*; D004492:Edetic Acid / Q000494:pharmacology; D006595:Hexosamines / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D008055:Lipids / Q000032:analysis; D008970:Molecular Weight; D010758:Phosphorus / Q000032:analysis
D001426:Bacterial Proteins; D002021:Buffers; D002241:Carbohydrates; D002254:Carbonates; D006595:Hexosamines; D008055:Lipids; D010758:Phosphorus; D004492:Edetic Acid