Forces causing osmotic pressure.

An analysis of forces acting on a simple system that includes a solvent, solute, and a semipermeable membrane shows the basic mechanism of osmotic pressure can be expressed as any of three forces: that due to the interaction of solute particles with the semipermeable membrane; that due to the cohesiveness of the solvent; or that due to the solute particles interacting with the free surface of the solvent.     

Porin activity in the osmotic shock fluid of Escherichia coli.

Osmotic shock fluid of Escherichia coli exhibited pore-forming activity. This activity could be followed by an in vitro assay based on the conductivity increase for ions due to the presence of pores in black lipid membranes. The histogram (the distribution of conductivity increments in a single pore experiment) obtained with osmotic shock fluid from E. coli was identical to the histogram obtained by detergent-solubilized porin isolated from the outer membrane. The osmotic shock fluid from porin-negative mutants also exhibited pore activity, although the histogram and ion specificity were different from those of porin. Antibodies raised against detergent-solubilized porin were able to form precipitin lines by the Ouchterlony immunodiffusion technique when shock fluids, but not detergent-solubilized porin, were used. These antibodies prevented the formation of pores when shock fluids contained porin but not when shock fluids obtained from porin-negative mutants were used. Macroscopic membrane conductivity of shock fluids due to porin exhibited a concentration dependence, in contrast to detergent-solubilized porin. These results indicate that the hydrodynamic properties of periplasmic or "soluble" porin are different from those of the detergent-solubilized porin of the outer membrane. Periplasmic porin comprises about 0.7% of total protein in the osmotic shock fluid.     

The kinetics of colloid osmotic hemolysis. II. Photohemolysis

Many of the known features of photohemolysis have been organized in a kinetic model that simulates the lytic time-course in a variety of conditions. The model combines Nernst-Planck flux principles, the osmotic equilibrium model of Freedman and Hoffman, equations relating illumination parameters to ion permeability, and an empirical relation between cell volume and lysis. Model simulations are compared with experiments showing the dependence of lysis kinetics on sensitizer concentration and on the osmotic content of the reaction medium. Additional experiments demonstrate that the inherent osmotic fragility of erythrocytes is not altered by illumination conditions that cause major delayed lysis 23 h later. The successful simulations support the hypothesis that photohemolysis is a colloid osmotic lysis occurring in cells behaving as imperfect osmometers.     

High osmotic stress behavior of hyaluronate and heparin.

Using polyethylene glycol and dextran as osmotic stressing agents, the concentrations of hyaluronate and heparin were measured as a function of osmotic pressure II over the range of 0.03 to nearly 50 atmospheres. The experimental results were analyzed in terms of the Donnan osmotic pressure, the virial expansion, and Flory's first neighbor interaction parameter. In addition, II was looked at as a function of the reciprocal cube root of the concentration, which represents an average intermonomer spacing at high concentrations. The decay lengths in the so-called hydration region were found to be around 2.6 A and negligibly salt dependent. In the electrostatically dominated region the decay lengths were found to be dependent on the ionic strength, but not simply so. The osmotic compressibilities were also calculated, and were compared to compressibility data of corneal stroma and articular cartilage. These latter compressibilities were close to those for the pure hyaluronate and heparin, strengthening the evidence that glycosaminoglycans (GAGs) are largely responsible for connective tissue compressibility. Higher compressibilities for previously reported GAG data is thought to be related to the protein content of those samples.     

The colloid osmotic pressures of invertebrate body fluids.

Colloid osmotic pressures of the body fluids of twenty invertebrate species were measured directly. The results, which are generally lower than predicted values for the same species, pertain to several physiological questions: (1) they do not quantitatively explain the frequently observed hyperosmoticity of body fluids in species believed to be osmoconformers, indicating that the condition cannot be merely a consequence of a Gibbs-Donnan equilibrium; (2) the excess of hydrostatic over colloid osmotic pressure is very small. This result supports the hypothesis that the oxygen transport function of bloods with extracellular haemocyanins and haem proteins is limited by their colligative properties; (3) the pressure relationships and the absence of colloid osmotic activity in urine indicates that filtration contributes to urine formation in several species.     

Metabolic engineering of plants for osmotic stress resistance.

Genes encoding critical steps in the synthesis of osmoprotectant compounds are now being expressed in transgenic plants. These plants generally accumulate low levels of osmoprotectants and have increased stress tolerance. The next priority is therefore to engineer greater osmoprotectant synthesis without detriment to the rest of metabolism. This will require manipulation of multiple genes, guided by thorough analysis of metabolite fluxes and pool sizes.     

Transient inhibition of translation initiation by osmotic stress.

Cells respond and adapt to changes in the environment. In this study, we examined the effect of environmental stresses on protein synthesis in the yeast Saccharomyces cerevisiae. We found that osmotic stress causes irreversible inhibition of methionine uptake, transient inhibition of uracil uptake, transient stimulation of glucose uptake, transient repression of ribosomal protein (RP) genes such as CYH2 and RPS27, and the transient inhibition of translation initiation. Rapid inhibition of translation initiation by osmotic stress requires a novel pathway, different from the amino acid-sensing pathway, the glucose-sensing pathway, and the TOR pathway. The Hog1 MAP kinase pathway is not involved in the inhibition of either methionine uptake or translation initiation but is required for the adaptation of translation initiation after inhibition and the repression of RP genes by osmotic stress. These results suggest that the transient inhibition of translation initiation occurs as a result of a combination of both acute inhibition of translation and the long-term activation of translation by the Hog1 pathway.     

Protein diffusion in the periplasm of E. coli under osmotic stress

The physical and mechanical properties of the cell envelope of Escherichia coli are poorly understood. We use fluorescence recovery after photobleaching to measure diffusion of periplasmic green fluorescent protein and probe the fluidity of the periplasm as a function of external osmotic conditions. For cells adapted to growth in complete medium at 0.14–1.02 Osm, the mean diffusion coefficient <D_peri> increases from 3.4 μm² s⁻¹ to 6.6 μm² s⁻¹ and the distribution of D_peri broadens as growth osmolality increases. This is consistent with a net gain of water by the periplasm, decreasing its biopolymer volume fraction. This supports a model in which the turgor pressure drops primarily across the thin peptidoglycan layer while the cell actively maintains osmotic balance between periplasm and cytoplasm, thus avoiding a substantial pressure differential across the cytoplasmic membrane. After sudden hyperosmotic shock (plasmolysis), the cytoplasm loses water as the periplasm gains water. Accordingly, <D_peri> increases threefold. The fluorescence recovery after photobleaching is complete and homogeneous in all cases, but in minimal medium, the periplasm is evidently thicker at the cell tips. For the relevant geometries, Brownian dynamics simulations in model cytoplasmic and periplasmic volumes provide analytical formulae for extraction of accurate diffusion coefficients from readily measurable quantities.