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Volume 140, Issue 12

Expression of the P.69 pertactin adhesin in : fate of the carboxy-terminal domain Free

Abstract

The mature pertactin protein (P.69) of can be isolated from the bacterial cell surface as a polypeptide with an apparent molecular mass of 69000 Da as determined by sodium dodecyl sulphate gel electrophoresis. However the open reading frame of , the pertactin gene, encodes a polypeptide with a predicted molecular mass of 93478 Da, referred to as P.93. Expression of the gene in leads to the synthesis of the full-length P.93 polypeptide, which is rapidly processed to the mature P.69 protein located at the cell surface. The P.93 precursor polypeptide is processed at both termini. A 34 amino acid long signal sequence is removed from the amino-terminus and a polypeptide sequence of about 30000 Da (P.30) is cleaved from the carboxy-terminus. Deletion of the 3′ region of , encoding P.30, results in the expression of an intracellular form of P.69. Antiserum which recognizes P.30 was raised using synthetic peptides based on the primary amino acid sequence of the region. This anti-P.30 serum was used in a Western blot analysis of fractionated cells of and harbouring the intact gene. The P.30 polypeptide was readily detected in outer membrane fractions prepared from both of these bacterial species, although it could not be shown to be exposed at the cell surface.

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Keyword(s): adhesin , Bordetella pertussis , pertactin and secretion
© Society for General Microbiology, 1994
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1994-12-01
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Expression of the Bordetella pertussisP.69 pertactin adhesin in Escherichia coli: fate of the carboxy-terminal domain
Microbiology 140, 3301 (1994); https://doi.org/10.1099/13500872-140-12-3301
/content/journal/micro/10.1099/13500872-140-12-3301
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