A glyceraldehyde-3-phosphate dehydrogenase homolog in Borrelia burgdorferi and Borrelia hermsii.
A polyreactive monoclonal antibody recognized a 38.5-kDa polypeptide with amino-terminal sequence identity to conserved regions of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in Borrelia burgdorferi, the Lyme disease agent, and Borrelia hermsii, an agent of American relapsing fever. This monoclonal antibody also recognized GAPDH from other pathogenic spirochetes and other prokaryotes and eukaryotes as well. GAPDH activity was detected in sonicates of both B. burgdorferi and B. hermsii but not in live, intact organisms, indicating the possibility of a subsurface localization for the Borrelia GAPDH activity. Degenerate primers constructed from highly conserved regions of gapdh of other prokaryotes successfully amplified this gene homolog in both B. burgdorferi and B. hermsii. Nuclei acid and deduced amino acid sequence analysis of the 838-bp probes for each borrelia indicated 93.9% identity between B. burgdorferi and B. hermsii at the amino acid level. Amino acid identities of B. burgdorferi and B. hermsii with Bacillus stearothermophilus were 59.2% and 58.8% respectively. Southern hybridization studies indicated that the gene encoding GAPDH is located on the chromosome of each borrella. In other bacterial species, GAPDH has other functions in addition to its traditional enzymatic role in the glycolytic pathway. GAPDH may play a similar role in borrelias.
Anda
P
P
Centro Nacional de Microbiologia, Virologia e Immunologia Sanitarias, Instituto de Salud Carlos III, Madrid, Spain.
Gebbia
J A
JA
Backenson
P B
PB
Coleman
J L
JL
Benach
J L
JL
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000276:immunology; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000276:immunology; D001483:Base Sequence; D001898:Borrelia / Q000201:enzymology / Q000235:genetics* / Q000276:immunology; D015748:Borrelia burgdorferi Group / Q000201:enzymology / Q000235:genetics / Q000276:immunology; D002876:Chromosomes, Bacterial; D003429:Cross Reactions; D015342:DNA Probes; D005798:Genes, Bacterial; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D008193:Lyme Disease / Q000382:microbiology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012061:Relapsing Fever / Q000382:microbiology; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D015342:DNA Probes; D007075:Immunoglobulin M; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases