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Protonmotive force driven 6-deoxyglucose uptake by the oral pathogen, Streptococcus mutans Ingbritt

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Abstract

Streptococcus mutans Ingbritt was grown in glucose-excess continuous culture to repress the glucose phosphoenolpyruvate phosphotransferase system (PTS) and allow investigation of the alternative glucose process using the non-PTS substrate, (3H) 6-deoxyglucose. After correcting for non-specific adsorption to inactivated cells, the radiolabelled glucose analogue was found to be concentrated approximately 4.3-fold intracellularly by bacteria incubated in 100 mM Tris-citrate buffer, pH 7.0. Mercaptoethanol or KCl enhanced 6-deoxyglucose uptake, enabling it to be concentrated internally by at least 8-fold, but NaCl was inhibitory to its transport. Initial uptake was antagonised by glucose but not 2-deoxyglucose. Evidence that 6-deoxyglucose transport was driven by protonmotive force (Δp) was obtained by inhibiting its uptake with the protonophores, 2,4-dinitrophenol, carbonylcyanide m-chlorophenylhydrazine, gramicidin and nigericin, and the electrical potential difference (ΔΨ) dissipator, KSCN. The membrane ATPase inhibitor, N,N1-dicyclohexyl carbodiimide, also reduced 6-deoxyglucose uptake as did 100 mM lactate. In combination, these two inhibitors completely abolished 6-deoxyglucose transport. This suggests that the driving force for 6-deoxyglucose uptake is electrogenic, involving both the transmembrane pH gradient (ΔpH) and ΔΨ. ATP hydrolysis, catalysed by the ATPase, and lactate excretion might be important contributors to ΔpH.

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Abbreviations

DNP:

2,4-dinitrophenol

CCCP:

carbonylcyanide m-chlorophenylhydrazone

DCCD:

N,N1-dicyclohyxyl carbodiimide

Δp:

protonmotive force

ΔpH:

transmembrane pH gradient

ΔΨ:

transmembrane electrical potential difference

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Authors and Affiliations

  1. PHLS Centre for Applied Microbiology and Research, Pathogenic Microbes Research Laboratory, Porton Down, SP4 OJG, Salisbury, Wilts, UK

    C. W. Keevil, A. S. McDermid, P. D. Marsh & D. C. Ellwood

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  1. C. W. Keevil
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  2. A. S. McDermid
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  3. P. D. Marsh
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  4. D. C. Ellwood
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Keevil, C.W., McDermid, A.S., Marsh, P.D. et al. Protonmotive force driven 6-deoxyglucose uptake by the oral pathogen, Streptococcus mutans Ingbritt. Arch. Microbiol. 146, 118–124 (1986). https://doi.org/10.1007/BF00402337

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  • DOI: https://doi.org/10.1007/BF00402337

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