Modeling the Role of Negative Cooperativity in Metabolic Regulation and Homeostasis

A significant proportion of enzymes display cooperativity in binding ligand molecules, and such effects have an important impact on metabolic regulation. This is easiest to understand in the case of positive cooperativity. Sharp responses to changes in metabolite concentrations can allow organisms to better respond to environmental changes and maintain metabolic homeostasis. However, despite the fact that negative cooperativity is almost as common as positive, it has been harder to imagine what advantages it provides. Here we use computational models to explore the utility of negative cooperativity in one particular context: that of an inhibitor binding to an enzyme. We identify several factors which may contribute, and show that acting together they can make negative cooperativity advantageous.     

Isolation and characterization of the plasma membrane from the yeast Pichia pastoris

Despite similarities of cellular membranes in all eukaryotes, every compartment displays characteristic and often unique features which are important for the functions of the specific organelles. In the present study, we biochemically characterized the plasma membrane of the methylotrophic yeast Pichia pastoris with emphasis on the lipids which form the matrix of this compartment. Prerequisite for this effort was the design of a standardized and reliable isolation protocol of the plasma membrane at high purity. Analysis of isolated plasma membrane samples from P. pastoris revealed an increase of phosphatidylserine and a decrease of phosphatidylcholine compared to bulk membranes. The amount of saturated fatty acids in the plasma membrane was higher than in total cell extracts. Ergosterol, the final product of the yeast sterol biosynthetic pathway, was found to be enriched in plasma membrane fractions, although markedly lower than in Saccharomyces cerevisiae. A further characteristic feature of the plasma membrane from P. pastoris was the enrichment of inositol phosphorylceramides over neutral sphingolipids, which accumulated in internal membranes. The detailed analysis of the P. pastoris plasma membrane is discussed in the light of cell biological features of this microorganism especially as a microbial cell factory for heterologous protein production.     

Abiotic edaphic factors affecting the growth of a threatened North American Sunflower, Helianthus paradoxus (Asteraceae)

This study evaluated the relationships among soil moisture, soil salinity, and soil oxygen on the growth of Helianthus paradoxus (Asteraceae), a threatened inland salt marsh species of western North America. The study was conducted in large growth boxes (1×2×0.3 m) tilted at an angle to achieve a saturated to dry water gradient similar to that found in the marsh. This experimental design allowed the evaluation of major abiotic factors (soil moisture and soil salinity) which have been shown to be potentially important for this species, while removing major biotic factors, such as competition from other community dominants. Maximum aboveground biomass occurred in the middle rows of the boxes, where surface soil water was reduced and subsurface soil water was intermediate in the gradient. Regression analyses indicated that H. paradoxus would grow best where surface soil water is approximately 5%, subsurface soil water ranges from 20 to 30%, and where surface soil salinity is less than 0.5 g kg⁻¹. Edaphic variables, particularly soil moisture and soil salinity, affect the growth of H. paradoxus. Data presented here suggest that the survival of this species depends on maintenance of the hydrologic regime.     

Conformations of blood group H-active oligosaccharides of ovarian cyst mucins

Spectroscopic data and conformational energy calculations are reported for eight oligosaccharides from ovarian cyst mucins and from human milk, the nonreducing terminals of which have fucose (α1 → 2)galactose linked either (β1 → 3) (type I) or (β1 → 4)(type II) to N-acetylglucosamine or in (β1 → 3) linkage to galactosaminitol. The fully assigned proton nmr spectra are reported along with nuclear Overhauser enhancement (NOE) data. Amide proton coupling constants and vacuum-uv CD spectra provide information on the amide plane orientation and amide environment. Our results imply that the fucosidic dihedral angles are similar for all three cases and that the substantial differences in the chemical shifts of the fucosyl protons of type I, type II, and 3-ol chains result from different perturbations by the amide group of the residue to which the β-galactose is linked. Stereopair diagrams of conformational models for both type I and II H chains are presented that are consistent with NOE, coupling constants, conformational energy calculations, and the CD data. While the temperature dependence of the observed NOE of penta- and hexasaccharides indicates that their rotational correlation times are strongly temperature dependent, we conclude that the conformations are essentially independent of temperature.     

Darting and marking techniques for an arboreal forest monkey,Cercopithecus ascanius

Techniques are described for capturing and marking redtail monkeys (Cercopithecus ascanius). An air-powered darting rifle and syringe darts loaded with a Ketamine-Rompun mixture were used for capture. Ketamine was used for maintaining anesthesia. Monkeys were darted 48 times and captured 27 times. In 24 of the 27 captures, the monkeys were released unharmed. Adult males were marked with radiotransmitters attached to collars of nylon webbing. Females received nylon webbing collars with colorcoded plastic washers for identification.