Flagellar preparations from Pseudomonas aeruginosa: isolation and characterization

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Abstract
Flagella from various strains of P. aeruginosa were isolated by shearing the flagella followed by differential centrifugation to obtain typical filaments as viewed through an EM. Electrophoretic analysis showed a major protein band corresponding to a flagellin with a MW of 53,000. Among the strains tested, flagellar antigen (FAg) preparations isolated from strains 1244 and 1210 routinely gave the highest percentage of flagellin, with the least amount of protein impurities, when grown on succinate-mineral salts medium. All FAg preparations contained 3-10 .mu.g of 2-keto-3-deoxyoctonate-positive material/mg of protein. Strain PA-103 lacked flagella and exhibited no flagellin band; preparations from PA-103 had a relatively higher content of 2-keto-3-deoxyoctonate. The isolation of a highly purified, single-banded flagellin could be accomplished by elution of the 53,000 MW gel band. Amino acid analysis showed 16 amino acids, but no proline. Antisera to FAg preparations were used to demonstrate inhibition of motility of strains RM-46 and M-2. Heated RM-46 FAg antisera (mouse) and PA-103 antisera did not inhibit motility.