Phosphorus nuclear magnetic resonance of Acholeplasma laidlawii cell membranes and derived liposomes.

1. The 129 MHz 31P-NMR spectrum of Acholeplasma laidlawii membranes is very similar to the spectrum of the derived liposomes and is a typical "solid state" spectrum in which the major contribution to the linewidth is made by the chemical shift anisotropy. From the value of the chemical shift anisotropy an order parameter of 0.15 is estimated for the lipid phosphates in both membranes. 2. The 31P-NMR spectrum of the A. laidlawii membrane is insensitive to pronase digestion of 4-60% of the membrane proteins and subsequent cytochrome C binding. These results indicate that either no strong lipid polar headgroup-protein interactions occur in the membrane or that the lipid-protein "complexes" in the membrane have a fast rotation (Tc shorter than 10(-6)S) along an axis perpendicular to the plane of the membrane. 3. Phospholipase A2 degrades all the phosphatidylglycerol in the membrane. The resulting membrane contains a phosphoglycolipid as the sole phosphorus-containing compound. The 31P-NMR spectrum of these membranes is identical to the spectrum of the native membranes suggesting a similar motion for the phosphate groups in both lipids. 4. Ca2+ binding to liposomes prepared from either the total polar lipids or the total phosphorus-containing lipids isolated from the A. laidlawii membrane does not affect the 21P-NMR spectrum. 5. The 31P-NMR spectrum of the membranes and derived liposomes, however, is sensitive to lipid phase transitions. When the membrane lipids are in the gel state a broadening of the 31P resonance occurs demonstrating that the polar head group motion in a biological membrane is more restricted below the lipid-phase transition temperature.     

Purification,properties, and mode of action of hemicellulase I produced by Ceratocystis paradoxa

A culture isolate (CP₂) of the fungal plant pathogen Ceratocystis paradoxa produces at least five extra-cellular hemicellulases when grown on a medium containing a commercial hemicellulose as inducer. One of the five enzymes, hemicellulase I (HC-I), was purified by ammonium sulphate precipitation, ion-exchange chromatography (DEAE-Sephadex and then Cellex-CM), and iso-electric focusing at pH 3–10 and 8–10. HC-I behaves as a single protein on electrophoresis at pH 6.0 and 8.4. The enzyme degrades hemicellulose B (an arabino-4-O-methylglucuronoxylan) and arabinoxylan to arabinose, xylose, xylobiose (Xyl₂; β-D-Xylp-(1→4)-D-Xyl), and a mixture of arabinose-xylose and xylose oligosaccharides (AraXyl_n and Xyl_n, where n  3, 4, or 5). The enzyme is deduced to be an endo-enzyme. Xylotetraose (Xyl₄) was the lowest homologue of the xylose oligosaccharides attacked, yielding xylobiose and xylotriose (Xyl₃) only. A mechanism is postulated for this reaction. AraXyl₂AraXyl₅ were slowly hydrolysed to arabinose and the respective xylose saccharide (Xyl₂Xyl₅), and thence to Xyl₂ and Xyl₃. Hydrolysis of the arabinofuranosyl linkage probably does not occur at the same active site as for the xylose oligosaccharides. Hemicellulose B fractions from different sources appeared to be degraded by HC-I. The enzyme showed optimum activity at pH 5.5 and 40°, and K_m was 4.24 mg of hemicellulose/ml.     

Structures of the oligosaccharides from the enzymic hydrolysis of hemicellulose by a hemicellulase of Ceratocystis paradoxa.

And endo-hemicellulase (HC-II) of Ceratocystis paradoxa degraded spear-grass hemicellulose B to a series of mixed oligosaccharides. Four neutral oligosaccharides (AraXyl2, AraXyl3, Xyl2, and Xyl3), isolated by preparative paper chromatography, were shown by enzymic and methylation techniques to constitute a series of beta-(1 leats to 4)-D-xylose and O-alpha-L-arabinofuranosyl-(1 leads to 3)-O-beta-D-xylopyranosyl-(1 leads to 4)-O-beta-D-xylopyranosyl-(1 leads to 4)-D-xylose, respectively, the latter being a new compound.     

Photochemical labeling of the cytoplasmic surface of the membranes of intact human erythrocytes.

N-(4-azido-2-nitrophenyl)-2-aminoethyl sulfonate (NAP-taurine), a photolabile nitrene precursor, has been shown to permeate the human erythrocyte membrane at 37degrees but not at 0 degrees. Utilizing this differential permeability, we have loaded intact erythrocytes with NAP-[35S]taurine in the dark at 37degrees, cooled them to 0 degrees, washed them free of external NAP-[35S]taurine in the dark and cold, and photolyzed them, resulting in labeling of hemoglobin and of proteins on the cytoplasmic surface of the membrane. These experiments complement those previously reported on the labeling of the external surface of the membranes with this reagent (Staros, J. V., and Richards, F. M. (1974) Biochemistry 13, 2720-2726).     

The ultrastructure of the small granular, proteinaceous epidermal cells of some British lumbricids (Annelida) and a reassessment of the identity of the so-called albumen cells

The ultrastructure of the small granular, proteinaceous cells of 11 species of lumbricid earthworm is described and no species differences were recorded. The cells are characterized by the presence of membrane-bound, electron dense granules (0.6–0.7 urn in diameter) arising from polarized Golgi systems in close topographical relationship to the granular endoplasmic reticulum. Variations in electron density of the granules appear to be associated with maturation of the granules. The granules show a finely reticular substructure at high magnification. A less than fully mature stage of this cell type is described.
The occurrence of the small granular cell type in the annelids is discussed, as is the possible function of its secretion.
The confusions in the lumbricid literature concerning this cell type are discussed, and the much referred to figure of the 'albumen' cell type in English texts is shown not to be equivalent to the small granular, proteinaceous type referred to in this and a previous histochemical study.     

Insights into the feeding ecology of the Seychelles Black Parrot Coracopsis barklyi using two monitoring approaches

Feeding ecology is an important factor for the survival of a species and knowledge of its parameters is a prerequisite for successful conservation work. In this study we describe the feeding ecology of the endemic Seychelles Black Parrot Coracopsis barklyi on Praslin, Seychelles, the only island on which this parrot is resident. We compared two methods to evaluate feeding choices: incidental observations and feeding walks on 25 transects in all habitat types. Black parrots fed on 46 different species, bringing the total number of known food plants to 53 species. They predominantly consumed endemic and native species (58% of observed feeding bouts), mainly their fruit pulp (in 68% of feeding bouts), followed by buds (15%) and seeds (37%) with occasional observations of leaves, bark and scale insects. The incidental method rendered many more observed bouts than the transect approach and the ratios of consumed species differed between methods but the transect results are regarded as more representative. The incidental method is not suitable for quantitative conclusions but complements the transect method, providing information about rarely occurring feeding events.     

Genetic Screens Identify Host Factors for SARS-CoV-2 and Common Cold Coronaviruses

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Conformational transitions in N-linked oligosaccharides

An assignment strategy involving 1H-1H correlated spectroscopy (COSY), relayed correlation spectroscopy (RECSY), nuclear Overhauser effect spectroscopy (NOESY), and triple quantum filtered correlated spectroscopy (TQCOSY) is described for six related N-linked oligosaccharides. These are of three "types", i.e., complex, bisected complex, and oligomannose. Using spin-spin coupling constant data derived from these assignments, together with semiempirical quantum mechanical energy calculations, we have examined the rotamer distributions at the Man alpha 1-6Man beta-linkage in each structure, and additionally at the Man alpha 1-6Man alpha-linkage in oligomannose oligosaccharides. We show that while several primary sequence differences are "passive", certain key residues modulate the orientation of the alpha 1-6 arms. These residues may be proximal or distal to the site of the conformational change. There is no direct correlation between these perturbations and the oligosaccharide type. These data are discussed in terms of the proposed recognition function of oligosaccharides in biological systems.