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Ion diffusion potentials across mycoplasma membranes determined by a novel method using a carbocyanine dye.

Abstract：

:The influence of transmembrane ion fluxes on mycoplasma membrane potentials was studied. Electric membrane potential was calibrated vs fluorescence intensity of a potential-sensitive carbocyanine dye according to delta psi = (RT/F) X log([aIN(1 - IN) - b]/Kint), where IN = I/I0, I0 = maximal fluorescence intensity (obtained for delta psi----infinity), and a and b are constants. Fluorescence intensity was calibrated vs membrane potential by inducing a K+ diffusion potential. The calibration procedure was based on the assumption that in the presence of valinomycin the membrane potential was determined entirely by K+ diffusion. Then the dependence of fluorescence intensity on the external K+ concentration, Kext, could be described by Ival = I0[1 + a/(Kext + b)]-1. For Mycoplasma mycoides subsp. capri and enterococci, the constants were determined from experimental data using nonlinear least-squares computer-assisted methods. The validity of our assumption was proved using the "null-point" method. Here the Ca2+ ionophore A23187 and varying external Ca2+ concentrations were used to change the membrane potential experimentally. K+ and Na+ diffusion potentials significantly contributed to mycoplasma membrane potential whereas Cl- had no influence. Under growth conditions the mycoplasma membrane potential was estimated to be delta psi = -68 mV.

journal_name

Arch Biochem Biophys

journal_title

Archives of biochemistry and biophysics

authors

Schummer U,Schiefer HG

doi

10.1016/0003-9861(86)90623-5

subject

Has Abstract

pub_date

1986-02-01 00:00:00

pages

553-62

issue

2

eissn

0003-9861

issn

1096-0384

pii

0003-9861(86)90623-5

journal_volume

244

pub_type

杂志文章

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