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Infection and Immunity, February 1999, p. 754-759, Vol. 67, No. 2
0019-9567/99/$00.00+0

Identification and Characterization of PtlC, an Essential Component of the Pertussis Toxin Secretion System

David M. Cook,dagger Karen M. Farizo, and Drusilla L. Burns*

Laboratory of Pertussis, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892

Received 19 June 1998/Returned for modification 10 September 1998/Accepted 5 November 1998

PtlC is a member of a set of proteins necessary for the secretion of pertussis toxin (PT) from Bordetella pertussis. Using polyclonal antibodies specific for PtlC, we identified PtlC as a protein with an apparent molecular weight of 85,000 that localizes to the membrane fraction of bacterial cell extracts. We found that a mutant strain of B. pertussis that contains an in-frame deletion in ptlC is unable to secrete PT and that PT secretion is fully restored by expressing ptlC in trans, indicating that PtlC is essential for transport of PT across the bacterial membrane(s). PT secretion was inhibited in wild-type B. pertussis after introduction of a plasmid expressing a mutant ptlC altered in the putative nucleotide-binding region, suggesting that this region of PtlC is essential for proper function. Moreover, the observed dominant negative phenotype suggests that PtlC either functions as a multimer or interacts with some other component(s) necessary for secretion of PT.


* Corresponding author. Mailing address: CBER/FDA, HFM-434, Building 29, Room 418, 8800 Rockville Pike, Bethesda, MD 20892. Phone: (301) 402-3553. Fax: (301) 402-2776. E-mail: burns{at}cber.fda.gov.

dagger Present address: MJ Research, Inc., 136 Coolidge Ave., Watertown, MA 02172.


Infection and Immunity, February 1999, p. 754-759, Vol. 67, No. 2
0019-9567/99/$00.00+0



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