Molecular and immunological characterization of a 64-kDa protein of Actinobacillus actinomycetemcomitans.
The 64-kDa protein to which about half the sera from patients with localized juvenile periodontitis and rapidly progressive periodontitis reacted strongly was purified from Actinobacillus actinomycetemcomitans Y4. Determination of the N-terminal sequence of the protein revealed that it was a GroEL-like protein. The DNA fragment containing the groEL gene of A. actinomycetemcomitans was amplified by polymerase chain reaction, and the groESL operon was cloned by using colony hybridization with the amplified fragment from A. actinomycetemcomitans chromosomal DNA. Sequence analysis revealed that structures of the operon and its products were typical in gram-negative bacteria. Rabbit polyclonal antibodies to the 64-kDa protein cross-reacted with approximately 65-kDa proteins of Haemophilus aphrophilus, Haemophilus influenzae, Haemophilus paraphrophilus, Escherichia coli and Eikenella corrodens but not with any cellular proteins of Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum. It is possible that antibodies reactive to the 64-kDa protein in periodontitis patients are induced by the cross-reactivity with the hsp60 proteins of other bacteria.
Nakano
Y
Y
Department of Preventive Dentistry, Kyushu University Faculty of Dentistry, Fukuoka.
Inai
Y
Y
Yamashita
Y
Y
Nagaoka
S
S
Kusuzaki-Nagira
T
T
Nishihara
T
T
Okahashi
N
N
Koga
T
T
D016976:Aggregatibacter actinomycetemcomitans / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000276:immunology*; D000942:Antigens, Bacterial / Q000276:immunology; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D001438:Bacteroidaceae / Q000737:chemistry / Q000276:immunology; D001483:Base Sequence; D018835:Chaperonin 10 / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D018834:Chaperonin 60 / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D003429:Cross Reactions; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000737:chemistry; D005798:Genes, Bacterial; D006090:Gram-Negative Bacteria / Q000737:chemistry / Q000276:immunology; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009876:Operon; D010518:Periodontitis / Q000276:immunology / Q000382:microbiology; D015183:Restriction Mapping; D017421:Sequence Analysis; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D018835:Chaperonin 10; D018834:Chaperonin 60; D004269:DNA, Bacterial; D006360:Heat-Shock Proteins