Decorin-binding protein of Borrelia burgdorferi is encoded within a two- gene operon and is protective in the murine model of Lyme borreliosis

Kayla E. Hagman, Pekka Lahdenne, Taissla G. Popova, Stephen F. Porcella, Darrin R. Akins, Justin D. Radolf, Michael V. Norgard

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Abstract

Isolated outer membranes of Borrelia burgdorferi were used in immunoblotting experiments with sera from immune mice to identify new putative Lyme disease vaccine candidates. One immunoreactive polypeptide migrated on polyacrylamide gels just proximal to outer surface protein C and comigrated with [3H] palmitate-labeled polypeptides. A degenerate oligonucleotide primer based upon internal amino acid sequence information was used to detect the corresponding gene within a B. burgdorferi total genomic library. The relevant open reading frame (ORF) encoded a polypeptide comprised of a 24-amino-acid putative signal peptide terminated by LLISC, a probable consensus sequence for lipoprotein modification, and a mature protein of 163 amino acids. Immunoblots of a recombinant fusion protein corresponding to this ORF supported the idea that the encoded protein was a previously reported decorin-binding protein (DBP) of B. burgdorferi N40 (B. P. Guo, S. J. Norris, L. C. Rosenberg, and M. Hook, Infect. Immun. 63:3467- 3472, 1995). However, further DNA sequencing revealed the presence of a second ORF, designated ORF-1, whose termination codon was 119 bp upstream of the dbp gene. ORF-1 also encoded a putative lipoprotein with a mature length of 167 amino acids. Northern blots, Southern blots, and primer extension analyses indicated that ORF-1 and dbp comprised a two-gene operon located on the 49-kb linear plasmid. Both proteins, which were 40% identical and 56% similar, partitioned into Triton X-114 detergent extracts of B. burgdorferi isolated outer membranes. Mice infected with B. burgdorferi produced high titers of antibodies against the ORF-1-encoded protein and DBP during both early and later stages of chronic infection. Both DBP and the ORF-1-encoded protein were sensitive to proteinase K treatment of intact borreliae, suggesting that they were surface exposed. In active immunization experiments, 78% of mice immunized with recombinant DBP were immune to challenge. While it is not clear whether the two lipoproteins encoded by the ORF-1-dbp operon have analogous decorin-binding functions in vivo, the combined studies implicate DBP as a new candidate for a human Lyme disease vaccine.

Original languageEnglish (US)
Pages (from-to)2674-2683
Number of pages10
JournalInfection and Immunity
Volume66
Issue number6
StatePublished - Jun 1998

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Decorin
Borrelia burgdorferi
Lyme Disease
Operon
Open Reading Frames
Carrier Proteins
Genes
Lyme Disease Vaccines
Lipoproteins
Amino Acids
Peptides
Proteins
Recombinant Fusion Proteins
Borrelia
Endopeptidase K
DNA Primers
Membranes
Genomic Library
Terminator Codon
Palmitates

ASJC Scopus subject areas

  • Immunology

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Decorin-binding protein of Borrelia burgdorferi is encoded within a two- gene operon and is protective in the murine model of Lyme borreliosis. / Hagman, Kayla E.; Lahdenne, Pekka; Popova, Taissla G.; Porcella, Stephen F.; Akins, Darrin R.; Radolf, Justin D.; Norgard, Michael V.

In: Infection and Immunity, Vol. 66, No. 6, 06.1998, p. 2674-2683.

Research output: Contribution to journalArticle

Hagman, Kayla E. ; Lahdenne, Pekka ; Popova, Taissla G. ; Porcella, Stephen F. ; Akins, Darrin R. ; Radolf, Justin D. ; Norgard, Michael V. / Decorin-binding protein of Borrelia burgdorferi is encoded within a two- gene operon and is protective in the murine model of Lyme borreliosis. In: Infection and Immunity. 1998 ; Vol. 66, No. 6. pp. 2674-2683.
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abstract = "Isolated outer membranes of Borrelia burgdorferi were used in immunoblotting experiments with sera from immune mice to identify new putative Lyme disease vaccine candidates. One immunoreactive polypeptide migrated on polyacrylamide gels just proximal to outer surface protein C and comigrated with [3H] palmitate-labeled polypeptides. A degenerate oligonucleotide primer based upon internal amino acid sequence information was used to detect the corresponding gene within a B. burgdorferi total genomic library. The relevant open reading frame (ORF) encoded a polypeptide comprised of a 24-amino-acid putative signal peptide terminated by LLISC, a probable consensus sequence for lipoprotein modification, and a mature protein of 163 amino acids. Immunoblots of a recombinant fusion protein corresponding to this ORF supported the idea that the encoded protein was a previously reported decorin-binding protein (DBP) of B. burgdorferi N40 (B. P. Guo, S. J. Norris, L. C. Rosenberg, and M. Hook, Infect. Immun. 63:3467- 3472, 1995). However, further DNA sequencing revealed the presence of a second ORF, designated ORF-1, whose termination codon was 119 bp upstream of the dbp gene. ORF-1 also encoded a putative lipoprotein with a mature length of 167 amino acids. Northern blots, Southern blots, and primer extension analyses indicated that ORF-1 and dbp comprised a two-gene operon located on the 49-kb linear plasmid. Both proteins, which were 40{\%} identical and 56{\%} similar, partitioned into Triton X-114 detergent extracts of B. burgdorferi isolated outer membranes. Mice infected with B. burgdorferi produced high titers of antibodies against the ORF-1-encoded protein and DBP during both early and later stages of chronic infection. Both DBP and the ORF-1-encoded protein were sensitive to proteinase K treatment of intact borreliae, suggesting that they were surface exposed. In active immunization experiments, 78{\%} of mice immunized with recombinant DBP were immune to challenge. While it is not clear whether the two lipoproteins encoded by the ORF-1-dbp operon have analogous decorin-binding functions in vivo, the combined studies implicate DBP as a new candidate for a human Lyme disease vaccine.",
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AU - Norgard, Michael V.

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