Isolation of a corncob (coaggregation) receptor polypeptide from Fusobacterium nucleatum.
Corncobs, which are distinct morphological units formed by the ordered coaggregation of a filamentous microorganism and streptococci, can be made in vitro by using oral strains of Fusobacterium nucleatum and Streptococcus sanguis. Previous studies have shown that strains of F. nucleatum contain one of at least two different types of corncob receptor. The objective of this study was to isolate the receptor from F. nucleatum ATCC 10953 as the first step in the elucidation of the molecular basis of corncob formation. The cell envelope fraction from this bacterium was treated with trypsin, delipidated with chloroform-methanol, and subjected to ion-exchange chromatography. A single polypeptide (apparent Mr, 39,500), which was eluted from the column with 0.5 M sodium chloride and extracted with dodecyltrimethylammonium bromide to remove contaminating lipopolysaccharide, inhibited corncob formation between strain ATCC 10953 and S. sanguis CC5A. Similarly derived cell fractions from either F. nucleatum FDC 364 or Fusobacterium necrophorum failed to effect coaggregation in the inhibition assay. Amino acid analysis of the polypeptide showed a moderately hydrophobic character (polarity index, 41) and 11% basic residues. Antiserum made against the purified polypeptide agglutinated F. nucleatum ATCC 10953, neutralized the ability of this bacterium to form corncobs, and agglutinated whole cells of S. sanguis CC5A that were precoated with the receptor polypeptide. The identification and isolation of this receptor should greatly enhance our ability to define some of the complex intergeneric coaggregation mechanisms that are thought to occur in the human oral cavity.
Kaufman
J
J
Department of Microbiology, School of Dental Medicine, University of Pennsylvania, Philadelphia 19104.
DiRienzo
J M
JM
D000818:Animals; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000276:immunology / Q000302:isolation & purification*; D001667:Binding, Competitive; D005673:Fusobacterium / Q000032:analysis* / Q000378:metabolism / Q000502:physiology; D007106:Immune Sera / Q000494:pharmacology; D008565:Membrane Proteins / Q000302:isolation & purification; D008970:Molecular Weight; D011817:Rabbits; D011940:Receptor Aggregation; D011971:Receptors, Immunologic / Q000302:isolation & purification*; D013298:Streptococcus sanguis / Q000032:analysis / Q000378:metabolism / Q000502:physiology
D001426:Bacterial Proteins; D007106:Immune Sera; D008565:Membrane Proteins; D011971:Receptors, Immunologic