Reconstruction ofPasteurella multocidadermonecrotic toxin from three polypeptides

Autor: Katsumi Kume, Toyotsugu Nakai
Rok vydání: 1987
Předmět:
Zdroj: FEMS Microbiology Letters. 44:259-265
ISSN: 1574-6968
0378-1097
DOI: 10.1111/j.1574-6968.1987.tb02279.x
Popis: Whole Pasteurella multocida dermonecrotic toxin (DNT) molecule, indistinguishable from the native toxin in terms of molecular mass (Mr), electrophoretic behaviour, antigenicity, and toxicity, was reconstituted by mixing freshly purified fragment a (Mr, approx. 23000), fragment b (approx. 64000), and fragment c (approx. 74000) of P. multocida DNT after removal of urea and dithiothreitol by dialysis. The original dermonecrotizing activity was restored, when these dialyzed fragments were mixed in equimolar amounts based on Mr estimates, and this molar ratio gave the highest yield of toxic activity. The reconstitution of toxicity was not achieved in mixtures of any two of the three fragments. The dialyzed mixtures of fragments a and b (ab complex) and fragments b and c (bc complex) reassociated into protein components, and each of them migrated as a single protein band at the position corresponding to an Mr of approx. 87000 or approx. 138000 during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The dialyzed mixture of fragments a and c did not reassociate and migrated as two protein bands both at the positions corresponding to the Mrs of approx. 23000 (fragment a) and approx. 74000 (fragment c). The anti-‘fragment a’ and anti-‘ab complex’ antisera completely neutralized dermonecrotizing activity of the purified DNT, whereas the anti-‘fragment b’, anti-‘fragment c’, and anti-‘bc complex’ antisera did not.
Databáze: OpenAIRE
Externí odkaz: