Cloning, nucleotide sequence, and expression of a gene encoding an adhesin subunit protein of Helicobacter pylori

Journal of Bacteriology - Tập 175 Số 3 - Trang 674-683 - 1993
D. Gareth Evans1, T. Karjalainen1, David J. Evans1, David Y. Graham1, C H Lee1
1Department of Medicine, Baylor College of Medicine, Houston, Texas 77030.

Tóm tắt

Gene hpaA, which codes for the receptor-binding subunit of the N-acetylneuraminyllactose-binding fibrillar hemagglutinin (NLBH) of Helicobacter pylori, was cloned and sequenced. The protein expressed by hpaA, designated HpaA, was identified as the adhesin subunit on the basis of its fetuin-binding activity and its reactivity with a polyclonal, monospecific rabbit serum prepared against NLBH purified from H. pylori. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and Western blots (immunoblots) showed that the cloned adhesin has the same molecular weight (20,000) as that found on H. pylori. Also, HpaA contains a short sequence of amino acids (KRTIQK) which are all either identical or functionally similar to those which compose the sialic acid-binding motif of Escherichia coli SfaS, K99, and CFA/I. Affinity-purified antibody specific for a 12-residue synthetic peptide that included this sequence blocked the hemagglutinating activity of H. pylori and was shown by immuno-gold electron microscopy to react with almost transparent material on unstained H. pylori cells, which is consistent with previous observations concerning the location and morphology of the NLBH.

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Journal of Bacteriology

Tập 175 Số 3

674-683

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