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Temperature-induced membrane-lipid adaptation in Acanthamoeba castellanii.

ABSTRACT: A method has been developed for the separation of the major membrane fractions of Acanthamoeba castellanii after growth at different temperatures. The acyl-lipid compositions of individual membrane fractions, microsomal membranes, plasma membrane and mitochondria were analysed after a shift in culture temperature from 30 degrees C to 15 degrees C. The major change in lipid composition observed was an alteration in the relative proportions of oleate and linoleate. This reciprocal change was seen in all the membrane fractions, but occurred most rapidly in the phosphatidylcholine of the microsomal fraction. Thus, there appears to be a rapid induction of delta 12-desaturase activity in A. castellanii after a downward shift in growth temperature. Changes were also seen in the proportions of the n-6 C20 fatty acids, with a decrease in the proportions of icosadienoate and increases of icosatrienoate and arachidonate. However, unlike the alteration in oleate/linoleate ratios, this change was not seen in all the individual lipids of each membrane fraction.

SUBMITTER: Jones AL

PROVIDER: S-EPMC1132411 | biostudies-other | 1993 Feb

REPOSITORIES: biostudies-other

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Project description:The relationship between temperature-dependent changes in phagocytotic activity of Acanthamoeba castellanii and the fatty acid composition and physical properties of plasma membrane-enriched fractions were determined in cultures acclimated to 30 degrees C and 15 degrees C. Chilling (from 30 degrees C to 15 degrees C) had a very pronounced short-term inhibitory effect on phagocytosis only in stationary-phase cultures, which displayed a low degree of fatty acid unsaturation. A subsequent increase in phagocytosis by these cells was correlated with a low-temperature-induced increase in fatty acid unsaturation (shown previously [Jones, Lloyd and Harwood (1993) Biochem. J. 296, 183-188] to be due to n-6 desaturase induction). Plasma membrane-enriched fractions from 15 degrees C-acclimated cells also showed a marked increase in the relative proportion of polyunsaturated fatty acids. Steady-state fluorescence depolarization studies, using the membrane probe diphenylhexatriene, revealed increases in plasma membrane order with decreasing assay temperature. Over the upper assay-temperature range (25-40 degrees C), fluorescence anisotropy values were higher in membranes from 30 degrees C-acclimated cells; a 3.3 degrees C relative displacement of plots indicated that temperature-induced changes in membrane lipid composition compensated for approx. 22% of the ordering effect of low temperature. Changes in the temperature-dependence of fluorescence anisotropy, possibly corresponding to lateral phase separations or alterations in other bulk physical properties of membranes, occurred between 20 and 25 degrees C in membranes from 30 degrees C-acclimated cells and between 15 and 20 degrees C in membranes from 15 degrees C-acclimated cells. Fluorescence anisotropy plots were superimposed at assay temperatures between 5 and 15 degrees C. Short-term phagocytotic rates in whole cells decreased with assay temperature. Arrhenius discontinuities in rates of phagocytosis occurred at approx. 25.0 degrees C and 17.5 degrees C in 30 degrees C- and 15 degrees C-acclimated cells respectively, and in each case were thus within the temperature ranges of slope-change in the corresponding fluorescence anisotropy plots. The results show a direct correlation between plasma membrane fatty acid unsaturation, membrane physical properties and phagocytotic activity in A. castellanii. Therefore, a specific integrated physiological process has been correlated with fatty acid desaturase induction for the first time.

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Temperature-induced membrane-lipid adaptation in Acanthamoeba castellanii.

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