Interaction of the Bordetella pertussis filamentous hemagglutinin with heparin |
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Authors: | Franco D. Menozzi Claudie Gantiez Camille Locht |
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Affiliation: | Laboratoire de Microbiologie Génétique et Moléculaire, Institut Pasteur, Lille, France. |
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Abstract: | Heparin, a glycosaminoglycan synthesized in connective tissue-mast cells, appeared to inhibit the hemagglutination of rabbit erythrocytes induced by the filamentous hemagglutinin (FHA), a major adhesin of Bordetella pertussis. This inhibition suggested an interaction of heparin with the FHA region responsible for the hemagglutination activity. FHA-heparin interactions may play a role in bacterial attachment and persistence in the lungs during human pertussis. To confirm a direct FHA-heparin interaction, heparin was used as ligand in an affinity chromatography procedure. This technique allowed to purify FHA directly from the bacterial culture medium in a single-step using heparin-Sepharose CL-6B or Zetaffinity heparin 60 disks. The purified FHA was highly immunoreactive with anti-FHA monoclonal antibodies and showed no signs of degradation after 15 successive cycles of freezing-thawing. The described purification method is simple, and suitable for the rapid preparation of FHA. |
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Keywords: | Bordetella pertussis Hemagglutination Filamentous hemagglutinin Heparin Purification |
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