Probing the Borrelia burgdorferi surface lipoprotein secretion pathway using a conditionally folding protein domain.

Abstract

Surface lipoproteins of Borrelia spirochetes are important virulence determinants in the transmission and pathogenesis of Lyme disease and relapsing fever. To further define the conformational secretion requirements and to identify potential lipoprotein translocation intermediates associated with the bacterial outer membrane (OM), we generated constructs in which Borrelia burgdorferi outer surface lipoprotein A (OspA) was fused to calmodulin (CaM), a conserved eukaryotic protein undergoing calcium-dependent folding. Protein localization assays showed that constructs in which CaM was fused to full-length wild-type (wt) OspA or to an intact OspA N-terminal "tether" peptide retained their competence for OM translocation even in the presence of calcium. In contrast, constructs in which CaM was fused to truncated or mutant OspA N-terminal tether peptides were targeted to the periplasmic leaflet of the OM in the presence of calcium but could be flipped to the bacterial surface upon calcium chelation. This indicated that in the absence of an intact tether peptide, unfolding of the CaM moiety was required in order to facilitate OM traversal. Together, these data further support a periplasmic tether peptide-mediated mechanism to prevent premature folding of B. burgdorferi surface lipoproteins. The specific shift in the OM topology of sequence-identical lipopeptides due to a single-variable change in environmental conditions also indicates that surface-bound Borrelia lipoproteins can localize transiently to the periplasmic leaflet of the OM.

• Authors: Shiyong Chen, Wolfram R Zückert
• Journal: Journal of bacteriology (J Bacteriol) Vol. 193 Issue 23 Pg. 6724-32 (Dec 2011) ISSN: 1098-5530 [Electronic] United States
• PMID: 21965569 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Antigens, Surface
• Bacterial Outer Membrane Proteins
• Bacterial Secretion Systems
• Bacterial Vaccines
• Calmodulin
• Lipoproteins
• OspA protein

Topics

• Antigens, Surface (chemistry, genetics, metabolism)
• Bacterial Outer Membrane Proteins (chemistry, genetics, metabolism)
• Bacterial Secretion Systems
• Bacterial Vaccines (chemistry, genetics, metabolism)
• Borrelia burgdorferi (chemistry, genetics, metabolism)
• Calmodulin (genetics, metabolism)
• Humans
• Lipoproteins (chemistry, genetics, metabolism)
• Lyme Disease (metabolism, microbiology)
• Protein Folding
• Protein Structure, Tertiary
• Protein Transport