Calorimetric study of water in muscle tissue

Differential scanning calorimetry has been used to investigate the state of water in intact single muscle fibers of the giant barnacle, Balanus nubilus. The shapes of the melting curves suggest the presence of three types of water: unfrozen (or bound), free (or bulk) and intermediate water. The amount of unfrozen water per g protein was constant within experimental error. An increase in water content changed almost exclusively the amounts of free water. The amount of intermediate water varies only slightly with the fiber water content.     

Détermination des sites d'oxydation des dérivés du α-D-glucofuranose utilisés comme donneurs

The sites of oxidation, by catalytic transfer of H, of derivatives of 1,2-O-isopropylidene-α-D-glucofuranose suggest a regiospecific reaction. Compounds having vicinal hydroxyl groups at C-5 and C-6, or at C-3 and C-5, are oxidized at OH-5, whereas compounds having two hydroxyl groups at C-3 and C-6 or three hydroxyl groups give first aldehydes and then lactones.     

Evidence for uniformity of the carbohydrate chains in individual glycoprotein molecular variants

Pooled and alkylated α₁-acid glycoprotein was fractionated on a Con A-Sepharose column into two fractions : Con A-non reactive and Con A-reactive. The carbohydrate moiety from the α₁-acid glycoprotein Con A-reactive variant, obtained by hydrazinolysis and quantitative re-N-acetylation, contains only identical two-branched oligosaccharide chains. From the present work on α₁-acid glycoprotein and from previous studies on α₁-fetoprotein, one can assume that glycoprotein glycosylation occurs uniformly along each polypeptide chain giving it identical oligosaccharide units at each glycosylation site.     

Phosphorylation and dephosphorylation reactions by erythrocyte plasma membrane enzymes

Human erythrocyte membranes contain a phosphoprotein phosphatase able to dephosphorylate membrane protein previously phosphorylated by the endogenous protein kinase.The level of dephosphorylation obtained after prolonged incubation is about one half of the phosphorylated residues.The characteristics of this enzyme are similar to those described for the cytoplasmic phosphoprotein phosphatase.In a membrane preparation the phosphorylation and dephosphorylation reactions can be repeated, at least twice, achieving similar levels of phosphate esterified or hydrolyzed.The coordination of these two enzyme systems might play a role in some of the functions attributed to the protein kinase system.     

Sex-related differences in the expression of chick enterocyte butyrylcholinesterase during embryonic and post-hatching development

Summary The butyrylcholinesterase activity of chick enterocytes was studied from day 15 in ovo up to day 90 after hatching. The activities detected in both sexes at the level of jejuno-ileum change in a parallel manner, but the activity is always higher in the female than in the male during embryonic development. After hatching, the differences are less apparent although the study of the enzyme distribution along the intestine showed sex-related variations, mainly at the level of the anterior and middle parts of jejuno-ileum in the young adult. Analysis of butyrylcholinesterase by sucrose gradient centrifugation allowed to identify two globular soluble species (G₁ and G₄ forms). The G₄/(G₁ + G₄) ratio decreases during the development but this variation in the female does not parallel that observed in the male. Besides, the molecular form distribution along the intestine, studied after hatching, differs according to the sex. Taken together our results lead to hypothesize that the ontogeny and the regulation of the chick enterocyte butyrylcholinesterase depend on hormones.Abbreviations AChE Acetylcholinesterase - BuChE Butyrylcholinesterase     

Crystallization and prelilminary X-ray investigation of barster,the intracellular inhibitor of barnase

Crystals of barstar, the intracellular inhibitor of the extracellular ribonuclease produced by Bacillus amyloliquefaciens (barnase), were obtained through vapor phase equilibration using the hanging drop technique. Three crystal forms have been characterized. Forms I and II, crystallized eithr in potetragonal; they exhibit a superstructure along the c-axis. Form III crystals, suitable for a high resolution structure determination, were grown from 55-65% ammomnium sulfate. This crystal form is hexagonal and diffracts to at least 2 Å resolution at a synchrotron radiation source. It belongs to the hexagonal space group P6, with unit cell dimensions a = b = 143.6 Å, c = 35.6 Å. There are four molecules of barstar in the asymmetric unit. X-ray data have been collected to 2.2 Å Bragg sapcing. The structure determination is underway in order to analyze conformational changes of barstar upon complexation with barnas. © 1993 Wiley-Liss, Inc.     

Chromosomal R-banding with a monoclonal antidouble-stranded DNA antibody

Summary A monoclonal anti-DNA antibody (HB2) specific for poly dG-poly dC nucleotides was used to stain metaphasic lymphocyte or amniotic cell human chromosomes. HB2 fixation was revealed using either a peroxidase-or a rhodaminelabeled anti-mouse immunoglobulin antiserum. The staining pattern of the chromosomes was dependent on the HB2 concentration: R-banding could be observed at high antibody dilution. Previous trypsinization of metaphasic preparations demonstrated a precise and reproducible typical R-banding independent of the HB2 concentration. This technique appears to be an interesting alternative to other R-banding procedures. The specificity of the antibody allows a better understanding of the biochemical mechanism of R-banding.     

Effects of Germanium (Ge) on the silica spicules of the marine sponge Suberites domuncula: Transformation of spicule type

Germanium (Ge), in the form of germanic acid, at a Ge/Si molar ratio of 1.0 inhibits gemmule development and silica deposition in the marine demosponge Suberites domuncula. Lower Ge/Si ratios inhibit the growth in length of the silica spicules (tylostyles) producing short structures, but with relatively normal morphology and close to normal width; spherical protuberances occasionally occur on these spicules. A few of the short spicules possess completely round rather than pointed tips. Many of the latter develop when Ge is added (pulsed) to growing animals, thus inducing a change in spicule type. These results indicate that the growth in length of the axial filament is more sensitive to Ge inhibition than is silica deposition and that pointed spicule tips normally develop because the growth of the axial filament at the spicule tip is more rapid than silica deposition. Newly formed spicules initiate silica deposition at the spicule head but the absence of Ge-induced bulbs as in freshwater spicules (oxeas) leaves open the question of whether there is a silicification center(s) present in Suberites tylostyles. The morphogenesis of freshwater oxeas and of marine tyolstyles appears fundamentally different-bidirectional growth in the former and unidirectional growth in the latter. X-ray analysis demonstrate relatively uniform Ge incorporation into the silica spicules with considerable variation from spicule to spicule in the incorporated level. Increased silicic acid concentration induces the formation of siliceous spheres, suggesting that the axial filament becomes prematurely encased in silica.     

Glycan uniformity within molecular variants of transferrin with distinct affinity for concanavalin A

Human serum Transferrin has been fractionated on concanavalin A-Sepharose column into three fractions: Con A-non reactive (5 %), Con A-weakly reactive (30 %) and Con A-reactive (65 %). Each Con A-affinity transferrin variant contains a pair of identical oligosaccharide units either tri-branched in the Con A-non reactive variant or two-branched in the Con A-weakly reactive and Con A-reactive transferrin. This is an additional support to our proposal that glycosylation occurs uniformly along each polypeptide chain yielding identical oligosaccharide units at each glycosylated site.