On the three-dimensional structure and catalytic mechanism of triose phosphate isomerase

Triose phosphate isomerase is a dimeric enzyme of molecular mass 56 000 which catalyses the interconversion of dihydroxyacetone phosphate (DHAP) and D-glyceraldehyde-3-phosphate. The crystal structure of the enzyme from chicken muscle has been determined at a resolution of 2.5 A, and an independent determination of the structure of the yeast enzyme has just been completed at 3 A resolution. The conformation of the polypeptide chain is essentially identical in the two structures, and consists of an inner cylinder of eight strands of parallel beta-pleated sheet, with mostly helical segments connecting each strand. The active site is a pocket containing glutamic acid 165, which is believed to act as a base in the reaction. Crystallographic studies of the binding of DHAP to both the chicken and the yeast enzymes reveal a common mode of binding and suggest a mechanisms for catalysis involving polarization of the substrate carbonyl group.     

The production of ethanol by Saccharomyces cerevisiae immobilized in polycation-stabilized calcium alginate gels

Summary Polycation treatment of preformed calcium alginate beads produced a matrix with higher resistance to phosphate ions. The treatment of immobilized Saccharomyces cerevisiae in the calcium alginate beads inhibited respiration of the entrapped cells but did not reduce ethanol production.     

Estimate of the sticking probability for cells in uniform shear flow with adhesion caused by specific bonds

A theory is developed for the aggregation rate of cells in uniform shear flow when the cell-cell adhesion is mediated by bonds between specific molecules on the cell surfaces such as antigen and antibody or lectin and carbohydrate. The theory is based on estimates of the frequency and duration of cell-cell collisions and of the number of bonds formed and required to hold the cells together. For high shear rates, the sticking probability is a function of a single dimensionless parameter, A, that is proportional to G-2, with G the shear rate. For low shear rates, the sticking probability is a function of a second dimensionless parameter, A' proportional to G-1. In either case the rate of cell-cell sticking is a maximum when A (or A') congruent to 1.0. For small values of A (or A') the cells collide frequently, but do not stick, whereas for large values of A (or A') the cells collide infrequently, but stick with larger probability. Studies in Couette viscometer or other flow having approximately uniform shear can test these models.     

A circular dichroism study of human erythrocyte ghost proteins during exposure to 2450 MHz microwave radiation

The effect of 2450 MHz microwave radiation on the proteins of human erythrocyte ghosts has been investigated using circular dichroism spectroscopy. A specially constructed waveguide inserted into the spectropolarimeter allowed the continuous recording of optical activity before, during and after microwave irradiation. The data indicate that high levels of microwave radiation (600 mW/g, specific absorption rate) induce decreases in alpha-helical conformation that may result from both thermal vibrations and increased strain on the intramolecular hydrogen bonds that maintain secondary structure. The latter effect may result from differential intramolecular interactions with the oscillating electric field. Spectrin (bands 1 and 2) isolated from the ghosts was more sensitive to microwave irradiation than intact ghosts, and spectrin-depleted vesicles were the least sensitive. The data, therefore, indicate that the alpha-helical conformation of spectrin is altered by high levels of microwave radiation.     

Molecular complexes of amino acids with porphyrins as possible precursors of pigment-protein systems

The present communication gives experimental results of the studies of catalytic and photochemical properties of peptide-like compounds containing metalloporphyrins (hemoproteinoids and molecular complexes obtained through adsorption of porphyrins and amino acids on volcanic ash). The data suggest that molecular complexes of amino acids with porphyrins could have evolved in the course of chemical evolution and were intermediated between abiogenically synthesized molecules of amino acids and porphyrins and pigment-protein systems of living organisms.     

Elimination and replenishment of tricarboxylic acid-cycle intermediates in myocardium.

1. The contribution of Co2 fixation to the anaplerotic mechanisms in the myocardium was investigated in isolated perfused rat hearts. 2. K+-induced arrest of the heart was used to elicit a transition in the concentrations of the intermediates of the tricarboxylic acid cycle. 3. Incorporation of 14C from [14]bicarbonate into tricarboxylic acid-cycle intermediates was measured and the rates of the reactions of the cycle were estimated by means of a linear optimization program which solves the differential equations describing a simulation model of the tricarboxylic acid cycle and related reactions. 4. The results showed that the rate of CO2 fixation is dependent on the metabolic state of the myocardium. Upon a sudden diminution of cellular ATP consumption, the pool size of the tricarboxylic acid-cycle metabolites increased and the rate of label incorporation from [14C]bicarbonate into the cycle metabolites increased simultaneously. The computer model was necessary to separate the rapid equilibration between bicarbonate and some metabolites from the potentially anaplerotic reactions. The main route of anaplerosis during metabolite accumulation was through malate + oxaloacetate. Under steady-state conditions there was a constant net outward flow from the tricarboxylic acid cycle via the malate + oxaloacetate pool, with a concomitant anaplerotic flow from metabolites forming succinyl-CoA (3-carboxypropionyl-CoA).     

rme1 Mutation of Saccharomyces cerevisiae: map position and bypass of mating type locus control of sporulation.

Sporulation in Saccharomyces cerevisiae normally occurs only in MATa/MAT alpha diploids. We show that mutations in RME1 bypassed the requirements for both a and alpha mating type information in sporulation and therefore allowed MATa/MATa and MAT alpha/MAT alpha diploids to sporulate. RME1 was located on chromosome VII, between LEU1 and ADE6.     

Isolation and characterization of a unique protease from sporulating cells of Bacillus subtilis

Two proteases, designated I and II, have been isolated from sporulating cells of Bacillus subtilis. They were partially purified by ammonium sulfate fractionation, Sephadex chromatography and affinity columns. Protease I was found to be similar to an already characterized B. subtilis protease. Protease II is trypsin-like in its substrate specificity and is distinct from protease I in its pH optimum, pH stability, molecular weight, substrate specificity, heat stability and sensitivity to various inhibitors. While both enzymes were produced primarily during sporulation, they attained maximum levels of activity at different times. Distinct functions for these proteases in post exponential B. subtilis are likely.