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Volume 147, Issue 5

Insertion of fluorescent fatty acid probes into the outer membranes of the pathogenic spirochaetes and Free

Abstract

The authors examined the ability of octadecanoyl (C), hexadecanoyl (C) and dodecanoyl (C) fatty acid (FA) conjugates of 5-aminofluorescein (OAF, HAF and DAF, respectively) to insert into the outer membranes (OMs) of , and . Biophysical studies have demonstrated that these compounds stably insert into phospholipid bilayers with the acyl chain within the hydrophobic interior of the apical leaflet and the hydrophilic fluorescein moiety near the phospholipid head groups. Consistent with the known poor intrinsic permeability of the OM to hydrophobic compounds and surfactants, was not labelled with any of the FA probes. OAF inserted more readily into OMs of than into those of , although both organisms were completely labelled at concentrations at or below 2 μg ml. Intact spirochaetes were labelled with OAF but not with antibodies against known periplasmic antigens, thereby confirming that the probe interacted exclusively with the spirochaetal OMs. Separate experiments in which organisms were cooled to 4 °C (i.e. below the OM phase-transition temperatures) indicated that labelling with OAF was due to insertion of the probe into the OMs. , but not , was labelled by relatively high concentrations of HAF and DAF. Taken as a whole, these findings support the prediction that the lack of lipopolysaccharide renders and OMs markedly more permeable to lipophilic compounds than their Gram-negative bacterial counterparts. The data also raise the intriguing possibility that these two pathogenic spirochaetes obtain long-chain FAs, nutrients they are unable to synthesize, by direct permeation of their OMs.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): DAF, dodecanoylaminofluorescein , FA, fatty acid , fatty acid metabolism , flow cytometry , HAF, hexadecanoyl-aminofluorescein , membrane fluidity , MFI, mean fluorescence intensity , OAF, octadecanoylaminofluorescein , OM, outer membrane , OspA, outer surface protein A and TpCM, T. pallidum cultivation medium
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2001-05-01
2024-04-25
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Insertion of fluorescent fatty acid probes into the outer membranes of the pathogenic spirochaetes Treponema pallidum and Borrelia burgdorferi
Microbiology 147, 1161 (2001); https://doi.org/10.1099/00221287-147-5-1161
/content/journal/micro/10.1099/00221287-147-5-1161
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