Phase transitions of phospholipid vesicles under osmotic stress and in the presence of ethylene glycol.

The effects of poly(ethylene glycol) (PEG) on the phase transition of phospholipid multilamellar vesicles (MLVs) were investigated by using differential scanning calorimetry (DSC). Main transition temperature (Tm) and the pre-transition temperature (Tp) of neutral phospholipid-, DMPC-1, DPPC- and DSPC-MLVs increased with an increase in PEG concentration. The subtransition temperature of DPPC-MLV also increased with an increase in PEG concentration. These results could be qualitatively explained by enhancement of the lateral packing on the basis of the osmoelastic coupling theory. The pretransition temperature increased faster than the main transition temperature did with an increase in PEG concentration. The increment of Tm depended on the hydrocarbon chain length, the shorter the hydrocarbon chain length was, the larger the increment was. The transition width in the DSC peak was broadened with an increase in PEG concentration. These three above-mentioned effects are the main differences between the effects of the osmotic stress on the phase transition of MLVs and those of hydrostatic pressure. On the other hand, ethylene glycol (EG), which is the monomer of PEG, had a biphasic effect on transition temperature of DPPC-, DSPC-, and DMPC-MLV, reducing Tm and Tp at low concentrations, but increasing Tm and extinguishing pretransition at high concentrations. This is explained by the induction of an interdigitated gel phase at high concentrations of EG, which indicates that EG can easily penetrate into the head group region of the lipid, in contrast with PEG 6K, because EG is small. Temperature-EG concentration phase diagrams for the various PC-MLVs were determined.(ABSTRACT TRUNCATED AT 250 WORDS)     

How small can the difference among competitors be for coexistence to occur

Closely related competitors comprising ofEscherichia coli strains having the same metabolic system and differing only with a few bases on the glutamine synthetase gene in the plasmid pKGN were previously shown to coexist in a chemostat. The differences among these closely related competitors can be considered large enough to allow coexistence as the level of enzyme activity is different. To bring the difference among competitors to the slightest possible, the mutation was introduced on the noncoding region of the plasmid pKGN harbored in the wild-type strain (strain W). The new strain, strain W’, carries the plasmid pKGN’ with a 4-base insertion at theHind III site in the polycloning site of pKGN. As the noncoding region is a nucleotide segment that is not translated into amino acids, the relatedness between strains W and W’ is the closest possible from the genetic point of view. Interestingly, though both strains are almost identical, they can coexist stably in a chemostat irrespective of the initial population size. These experimental results suggest that in the natural ecosystem, no matter how akin competitors are, coexistence is not impossible.     

The VIG9 gene products from the human pathogenic fungi Candida albicans and Candida glabrata encode GDP-mannose pyrophosphorylase

We have identified two genomic DNA fragments from the human pathogenic fungi, Candida albicans (CaVIG9) and Candida glabrata (CgVIG9) that encode GDP-mannose pyrophosphorylase, a key enzyme for protein glycosylation. The VIG9 homologues of CaVIG9 and CgVIG9 complement an identified protein glycosylation-defective mutation, vig9, of Saccharomyces cerevisiae. The nucleotide sequences of the ORFs, which are 83 and 90% identical to that of the ScVIG9 protein, respectively, showed a predicted gene product homologous to S. cerevisiae GDP-mannose pyrophosphorylase. We examined the enzyme activity of a glutathione S-transferase fusion of each VIG9 gene to synthesize GDP mannose in the cell extracts of a heterologous Escherichia coli expression system. We also developed a method for detecting the enzyme activity using a non-radioactive substrate that would be applicable to high throughput screening.