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J Bacteriol, April 1998, p. 1618-1623, Vol. 180, No. 7
Departments of Microbiology and Molecular
Genetics and Medicine, University of California Irvine, Irvine,
California 92697,1 and
Department of
Microbiology, Umeå University, Umeå Sweden2
Received 6 August 1997/Accepted 30 December 1997
A model of the 66-kDa outer membrane protein (P66) of Lyme disease
Borrelia spp. predicts a surface-exposed loop near the C
terminus. This region contains an antigen commonly recognized by sera
from Lyme disease patients. In the present study, this region of P66
and homologous proteins of other Borrelia spp. were further
investigated by using monoclonal antibodies, epitope mapping of P66 of
Borrelia burgdorferi, and DNA sequencing. A monoclonal antibody specific for B. burgdorferi bound to the portion
of P66 that was accessible to proteolysis in situ. The linear epitope for the antibody was mapped within a variable segment of the
surface-exposed region. To further study this protein, the complete
gene of Borrelia hermsii for a protein homologous to P66
was cloned. The deduced protein was 589 amino acids in length and 58%
identical to P66 of B. burgdorferi. The B. hermsii P66 protein was predicted to have a surface-exposed
region in the same location as that of B. burgdorferi's
P66 protein. With primers designed on the basis of conserved sequences
and PCR, we identified and cloned the same regions of P66 proteins of
Borrelia turicatae, Borrelia parkeri, Borrelia coriaceae, and Borrelia anserina. The
deduced protein sequences from all species demonstrated two conserved
hydrophobic regions flanking a surface-exposed loop. The loop sequences
were highly variable between different Borrelia spp. in
both sequence and size, varying between 35 and 45 amino acids. Although
the actual function of P66 of Borrelia spp. is unknown, the
results suggest that its surface-exposed region is subject to selective pressure.
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
A Surface-Exposed Region of a Novel Outer Membrane
Protein (P66) of Borrelia spp. Is Variable in Size and
Sequence
*
Corresponding author. Mailing address: Department of
Microbiology and Molecular Genetics, University of California Irvine, Irvine, CA 92697-4025. Phone: (714) 824-5626. Fax: (714) 824-8598. E-mail: abarbour{at}uci.edu.
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