Deciphering Lipid Metabolism in Borrelia burgdorferito Better Understand Lyme Disease

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Lyme disease is the most common tick-born disease in the United States, with almost half a million cases diagnosed every year. Lyme disease is caused by the bacterium Borrelia burgdorferi (Bb), which is transmitted to humans when they are bitten by an infected tick.

In recent work, Hove et al from Colorado State University in Denver, USA, identified a potential galactosyltransferase, BB0572, in the Bb genome by searching the Bb genome for sequences similar to a known glycosyltransferase from a related bacterium.

The group demonstrated that the gene encoding BB0572 was actively expressed by the Bb bacterium. To prove BB0572 has galactosyltransferase activity, the group carried out assays with 14C-labeled cholesterol and extracts of Bb bacteria and of E coli engineered to overexpress the gene for the BB0572 protein. The products of the assay were separated using thin-layer chromatography (TLC), and the radioactive products detected on the TLC plates using a Sapphire Biomolecular Imager.

Since the release of this publication, the Azure Sapphire has been succeeded by the new Azure Sapphire FL, which was designed to be the flexible choice in bringing precise quantitation of nucleic acids and proteins. Learn more.

Many people with Lyme disease have antibodies to a cholesterol-based glycolipid, ACGal (produced by Bb), leading some scientists to propose ACGal as a potential Lyme disease vaccine candidate. How the bacterium produces ACGal, and the related CGal, has been unclear. This is because Bb cannot make its own cholesterol and must modify cholesterol from the host.

Western blot results

The results indicate CGal can be synthesized by BB0572. ACGal was not detected, possibly because the acyl donor was missing from the reaction. The identification of BB0572 as the enzyme likely responsible for the synthesis of CGal and ACGal opens up new areas of research including identifying how ACGal is synthesized from CGal and studying the importance of these lipids to both Bb physiology and the development of Lyme disease.

Fig 5A. TLC of lipids from cell free assays for the enzymatic incorporation of [26- 14C] cholesterol into cholesteryl-β-D-galacto-pyranoside (CGal).The formation of cholesteryl-β-D-galacto-pyranoside (CGal) (arrow) in whole cell lysates of Bb and recombinant E. coli expressing WT bb0572 (lane 1 and 2). As expected, CGal was not formed by the whole cell lysate of E. coli expressing mutant bb0572 ΔCFF/I/DGD (lane 3). Lane 4 is the empty expression vector control, and lane 6 and 7 are from boiled whole cell lysates of Bb and recombinant E. coli expressing WT bb0572, respectively. Note lane 5 was not used.

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Background on Borrelia burgdorferi

Bb does not produce any toxins, and the pathogenesis of Lyme disease in humans appears to be due to a strong immune response to the bacterium itself. The resulting long-lasting inflammatory response can damage several different organs including the brain, peripheral and central nervous systems, heart, and joints.

When Bb infection is recognized immediately, usually by a characteristic bullseye shaped rash around the site of the tick bite, Lyme disease can usually be cured by treatment with antibiotics. However, 20% to 30% of people with Lyme disease will not have the rash, and many will not know they are infected in time to be cured. This has led to a great interest in developing a vaccine to prevent Lyme disease.

In addition to phosphor imaging to detect radiolabeled targets, the Sapphire provides chemiluminescence, multi-channel fluorescence, NIR fluorescence, densitometry, and white light imaging of blots, gels, tissues, and more.

Learn more about the Sapphire Imager and how Azure can support your research by clicking here.

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