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Infection and Immunity, June 1999, p. 2874-2883, Vol. 67, No. 6
Departments of Microbiology & Molecular
Genetics and Medicine, University of California Irvine, Irvine,
California 92697
Received 11 December 1998/Returned for modification 28 January
1999/Accepted 2 March 1999
The outer membrane of Borrelia burgdorferi, the Lyme
disease agent, contains lipoproteins anchored by their lipid moieties and integral proteins with membrane-spanning regions. We used the
techniques of in situ proteolysis, immunofluorescence, in vitro growth
inhibition, and cross-linking with formaldehyde to characterize
topological relationships between P66, an integral membrane protein,
and selected Osp lipoproteins of B. burgdorferi. Protease
treatment of intact spirochetes cleaved P66 and Osp proteins but not
the periplasmic flagellin or the BmpA protein of the cytoplasmic membrane. P66 of cells lacking OspA, OspB, and OspC was more
susceptible to trypsin cleavage than was P66 of cells with these Osp
proteins. A monoclonal antibody against the surface loop of P66 bound,
agglutinated, and inhibited the growth of viable spirochetes lacking
OspA, OspB, OspC, and OspD but not of the cells that expressed OspA,
OspC, and/or OspD. When cells were fixed, the antibody bound to cells that express OspD and OspC but still not to cells with OspA. The close
association of OspA and P66 was confirmed by the crosslinking of the
two proteins by formaldehyde. These results show that Osp proteins,
particularly OspA, limit the access of antibody or trypsin to the
surface loop region of P66. The proximity and possible contact between
P66 and OspA (or other Osp proteins) may hinder the effectiveness of
antibodies to what otherwise would be an appropriate vaccine target.
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Access of Antibody or Trypsin to an Integral Outer
Membrane Protein (P66) of Borrelia burgdorferi Is Hindered
by Osp Lipoproteins
*
Corresponding author. Mailing address: Department of
Microbiology & Molecular Genetics, University of California Irvine,
Irvine, CA 92697-4025. Phone: (949) 824-5626. Fax: (949) 824-8598. E-mail: abarbour{at}uci.edu.
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