Molecular and immunological characterization of a novel polymorphic lipoprotein of Borrelia burgdorferi.
We describe the cloning, expression, and molecular characterization of a novel polymorphic Borrelia burgdorferi lipoprotein recognized by monoclonal antibody LA7. Sequence analysis revealed an open reading frame encoding a 21,866-Da polypeptide (IpLA7). Comparison with other known proteins indicated sequence similarity between IpLA7 signal peptides and those of other prokaryotic lipoproteins, including the immunodominant B. burgdorferi outer surface proteins OspA, OspB, pC, and OspD. Both natural IpLA-7 and recombinant IpLA-7 could be biosynthetically labeled with [3H]palmitate. Upon solubilization of intact B. burgdorferi with the nonionic detergent Triton X-114, IpLA7 was extracted together with other lipoproteins into the detergent phase. Indirect immunolabeling studies indicated that the epitope recognized by monoclonal antibody LA7 is mainly located in the periplasmic space. Two-dimensional gel electrophoresis and immunoblotting confirmed the calculated acidic pI of 5.7 for IpLA-7. The LA7 gene was shown to be species specific and to be located on the linear chromosome of B. burgdorferi. The analysis of 40 individual spirochetal isolates on the basis of restriction fragment length polymorphisms revealed considerable genotypic heterogeneity of LA7 corresponding to that previously found for ospA. Native IpLA-7 and recombinant IpLA-7 were recognized by immune sera from infected mice as well as some human sera derived from infected but healthy donors and may thus prove useful as an additional marker for the serodiagnosis of Lyme disease.
Wallich
R
R
Angewandte Immunologie, Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Simon
M M
MM
Hofmann
H
H
Moter
S E
SE
Schaible
U E
UE
Kramer
M D
MD
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D015748:Borrelia burgdorferi Group / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D006801:Humans; D007526:Isoelectric Point; D008074:Lipoproteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D008193:Lyme Disease / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012150:Polymorphism, Restriction Fragment Length; D012995:Solubility
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D008074:Lipoproteins