The potent bone-resorbing mediator of Actinobacillus actinomycetemcomitans is homologous to the molecular chaperone GroEL.
Actinobacillus actinomycetemcomitans is a Gram-negative bacterium implicated in the pathology of localized juvenile periodontitis, a condition involving rapid destruction of alveolar bone. We have established that gentle extraction of this bacterium in saline releases a proteinaceous fraction (which we have termed surface-associated material [SAM] which has potent osteolytic activity in the murine calvarial bone resorption assay. Fractionation of the SAM has now revealed that activity is associated with a 62-kD protein. This bone-resorbing activity can be blocked by a monoclonal antibody (raised to the whole bacterium) that is claimed to recognize a protein homologous to the Escherichia coli molecular chaperone GroEL. Purification of this bone-resorbing protein to homogeneity has been achieved by a combination of anion exchange, gel filtration, and ATP-affinity chromatography and the NH2-terminal sequence shows > 95% homology to E. coli GroEL. This GroEL homologue is found in the SAM of A. actinomycetemcomitans but is not found in the osteolytically active SAM from other Gram-negative or Gram-positive bacteria. The GroEL protein from E. coli, but not from Mycobacterium tuberculosis and Mycobacterium leprae, also showed activity in the bone resorption assay. We believe this to be the first observation that a molecular chaperone has the capacity to stimulate the breakdown of connective tissue.
Kirby
A C
AC
Department of Microbiology, Eastman Dental Institute for Oral Health Care Sciences, University of London, United Kingdom.
Meghji
S
S
Nair
S P
SP
White
P
P
Reddi
K
K
Nishihara
T
T
Nakashima
K
K
Willis
A C
AC
Sim
R
R
Wilson
M
M
D016976:Aggregatibacter actinomycetemcomitans / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D001681:Biological Assay; D001862:Bone Resorption; D002478:Cells, Cultured; D018834:Chaperonin 60 / Q000737:chemistry*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008809:Mice, Inbred C3H; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D009166:Mycobacterium leprae / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012886:Skull / Q000166:cytology / Q000187:drug effects / Q000502:physiology
D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D018834:Chaperonin 60