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Infection and Immunity, December 1998, p. 5636-5642, Vol. 66, No. 12
Institute for Animal Health,
Received 11 June 1998/Returned for modification 24 July
1998/Accepted 17 September 1998
Pasteurella multocida toxin (PMT) is a potent mitogen
that also affects bone resorption. PMT acts intracellularly and is
therefore postulated to have several domains involved in different
aspects of its function. The toxin contains eight cysteine residues.
Mutants with individual substitutions for each of these residues were constructed, and the effects of these on the biological activity of the
toxin were determined by cultured-cell assays. Only the most C-terminal
of the eight cysteines (C1165) was essential for full activity,
although mutation of the cysteine residue at position 1159 caused a
slight but reproducible loss of potency. In animal challenge
experiments, mutant toxin (C1165S) was not toxic to piglets, even at
doses exceeding a lethal dose of active PMT 1,000-fold. The mutant and
wild-type toxins displayed identical purification characteristics,
similar susceptibility to proteolytic digestion, and circular dichroism
profiles, which indicated that no gross structural changes had taken
place. The function of the essential C1165 residue is not yet known,
although its most likely role is an enzymatic one at or near the
catalytic center of the toxin.
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Activity of the Mitogenic Pasteurella
multocida Toxin Requires an Essential C-Terminal Residue
*
Corresponding author. Mailing address: Oral
Microbiology, Guy's, King's and St. Thomas' Dental Institute, Floor
28, Guy's Hospital, London, SE1 9RT, United Kingdom. Phone: 44 (0) 171 955 2848. Fax: 44 (0) 171 955 2847. E-mail:
a.lax{at}umds.ac.uk.
Infection and Immunity, December 1998, p. 5636-5642, Vol. 66, No. 12
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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