Interaction of the Bordetella pertussis filamentous hemagglutinin with heparin.
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.
Menozzi
F D
FD
Laboratoire de Microbiologie Génétique et Moléculaire, Institut Pasteur, Lille, France.
Gantiez
C
C
Locht
C
C
D018829:Adhesins, Bacterial; D000818:Animals; D001886:Bordetella pertussis / Q000378:metabolism*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006386:Hemagglutination Tests; D006388:Hemagglutinins / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism*; D015151:Immunoblotting; D011817:Rabbits; D010566:Virulence Factors, Bordetella
D018829:Adhesins, Bacterial; D006388:Hemagglutinins; D010566:Virulence Factors, Bordetella; C065273:filamentous hemagglutinin adhesin, Bordetella pertussis; D006493:Heparin