Saccharomyces cerevisiae structural cell wall mannoprotein

A novel mannoprotein fraction with an average molecular weight of 180 000 has been isolated from Saccharomyces cerevisiae mnn9 mutant cell wall that was solubilized by beta-glucanase digestion. The same material could be extracted from purified wall fragments with 1% sodium dodecyl sulfate. The protein component, 12% by weight, is rich in proline, whereas the carbohydrate, mainly mannose, is about evenly distributed between asparagine and hydroxyamino acids. Endoglucosaminidase H digestion of the isolated mannoprotein reduced its average molecular weight to 150 000, but the mannoprotein, while still embedded in the cell wall, was inaccessible to the enzyme. Biosynthesis and translocation of the mannoprotein were investigated by following incorporation of [3H]proline into this fraction. In the presence of tunicamycin, both mnn9 and wild-type X2180 cells made a mannoprotein fraction with an average molecular weight of 140 000, whereas in the absence of the glycosylation inhibitor, the mnn9 mutant made material with a molecular weight of 180 000 and the mannoprotein made by wild-type cells was too large to penetrate the polyacrylamide gel. Although the cell wall mannoprotein was resistant to heat and proteolytic enzymes, attempts to isolate the carbohydrate-free component failed to yield any characteristic peptide material.     

Substitutions of hydrophobic amino acids reduce the amyloidogenicity of Alzheimer's disease beta A4 peptides.

The deposition of amyloid protein aggregates in brain is the main pathological feature of Alzheimer's disease. Their principal constituent is a peptide termed beta A4, which comprises up to 43 amino acid residues. It is highly insoluble under physiological conditions and aggregates into filaments that form very dense clusters in vivo and in vitro. Based on a beta A4 prototype sequence spanning residues 10 to 42 or 43, we have designed analogues in which hydrophobic amino acid residues in position 17 to 20 were substituted by more hydrophilic residues. Depending on the kind of newly introduced amino acids and their position within the sequence, the substitution of only two residues led to variants exhibiting a broad spectrum of different properties. Common to them was a reduced beta-sheet content after solubilization in water and in the solid state. Some of the variants showed significantly reduced amyloidogenicity: although still forming filaments, they did not aggregate into the highly condensed depositions that are typical for amyloid. In addition, they could be solubilized in 200 mM-NaCl and KCl. When mixed with beta A4 peptides bearing the natural sequence, two of the analogues could inhibit the formation of filaments in vitro. These results demonstrate that a well-preserved hydrophobic core around residues 17 to 20 of beta A4 is crucial for the formation of beta-sheet structure and the amyloid properties of beta A4. The introduction of structural alterations within this region may guide the development of reagents for the therapy of Alzheimer's disease.     

Parvalbumins from frog skeletal muscle (Rana temporaria L.). Isolation and characterization. Structural modifications associated with calcium binding.

Two parvalbumins have been isolated from the skeletal muscle of Rana temporaria L. Amino acid composition, tryptic peptide maps, isoelectric points, calcium content and ultraviolet as well as circular dichroism spectra have been determined. Investigation on antigenic properties revealed no antigenic determinants common to both components. The two protein molecules appear to belong to far related gene lineages. They are also different from the parvalbumins found in Rana esculenta muscle. Modifications of physical parameters, associated with calcium binding and dissociation are described. While antigenicity remained essentially unchanged, conformational changes were revealed by alterations of circular dichroism spectra.     

Packing of ribosomes in crystals from the lizard Lacerta sicula

The packing of ribosomes in the large crystalline sheets found in the lizard Lacerta sicula has been investigated by electron microscopy. The ribosomes in each of the two layers composing a sheet are organised as tetramers on a P4 space group lattice. The two layers face in opposite directions and tend to be related to one another crystallographically, generating a family of P422 crystals of different unit cell dimensions. The projected structure of one layer was determined from negatively stained, isolated sheets by separating the contributions from each layer in Fourier transforms computed from electron micrographs. Comparison of the projection map with other, low resolution, analyses of images of isolated eukaryotic ribosomes indicates that the large subunit- small subunit axis lies approximately parallel to the plane of the sheet.     

Effect of hyperthermia on growth of Ehrlich ascites tumor cells

Hyperthermic treatment at 43 degrees C suppressed the growth of Ehrlich ascites tumor (EAT) cells in vitro. Incubation of EAT cells at 43 degrees C for as little as 1.5 h totally abolished the transplantability of the tumor. At the same time, the rate of cellular glucose uptake, the density of glucose transporter on the cells as well as the extent of thymidine, uridine and leucine incorporation were significantly reduced.     

Seasonal Changes of the Metabolites in the Leaves, Bark and Xylem Tissues of Olive Tree (Olea europaea. L) II. Carbohydrates

Seasonal changes of starch and soluble carbohydrates in leaves,bark and xylem tissues of olive tree were examined during acomplete annual cycle. Leaf starch and soluble carbohydrateswere detected at high levels during the spring and autumn metabolicallyactive periods. The low level of leaf starch in combinationwith the drastic reduction of soluble carbohydrates and mannitol,defined the summer period of the low metabolic state of thetree. The low leaf starch level in conjunction with the risensoluble carbohydrate levels in leaves in winter were associatedwith cold acclimation processes. The bark and xylem tissueswere performing as starch deposition sites, and differencesin the extent of starch accumulation in these tissues were detectedduring the seasons. The starch fluctuations in bark and xylemwere discussed in relation to the translocation of metabolitesand other physiological processes. Mannitol, the most abundantleaf carbohydrate, was examined in relation to the reducingsugars exported to the bark. The bark mannitol was examinedin conjunction with the sucrose, glucose and starch levels inthe maturing bark tissues and was correlated to the low wintertemperatures. During the winter there was a drastic reductionof mannitol circulation from the bark to xylem. Olive tree, Olea europaea, L, carbohydrates     

Entry of [(1,2-13C2)acetyl]-L-carnitine in liver tricarboxylic acid cycle and lipogenesis: a study by 13C NMR spectroscopy in conscious, freely moving rats.

The biochemical pathways involved in acetyl-L-carnitine utilization were investigated in conscious, freely moving rats by 13C NMR spectroscopy. Following 4-h [(1,2-13C2)acetyl]-L-carnitine infusion in fasted animals, the free carnitine levels in serum were increased, and an efflux of unlabelled acetyl-L-carnitine from tissues was observed. [(1,2-13C2)Acetyl]-L-carnitine was found to enter biosynthetic pathways in liver, and the acetyl moiety was incorporated into both cholesterol and 3-hydroxybutyrate carbon skeleton. In accord with the entry of [(1,2-13C2)acetyl]-L-carnitine in the mitochondrial acetylCoA pool associated with tricarboxylic acid cycle, the 13C label was also found in liver glutamate, glutamine, and glutathione. The analysis of the 13C-labelling pattern in 3-hydroxybutyrate and cholesterol carbon skeleton provided evidence that the acetyl-L-carnitine-derived acetylCoA pool used for ketone bodies synthesis in mitochondria was homogeneous, whereas cholesterol was synthesized from two different acetylCoA pools located in the extra- and intramitochondrial compartment, respectively. Furthermore, cholesterol molecules were shown to be preferentially synthesized by the metabolic route involving the direct channelling of CoA-activated mitochondria-derived ketone bodies into 3-hydroxy-3-methylglutarylCoA pathway, prior to equilibration of their acyl groups with extramitochondrial acetylCoA pool via acetoacetylCoA thiolase.