RESUMO
Mass spectrometric and NMR spectroscopic analyses of bound sialic acids from the starfish Asterias rubens revealed the presence of N-acetylneuraminic acid (4%), N-acetyl-8-O-methylneuraminic acid (12%), N-acetyl-9-O-acetyl-8-O-methylneuraminic acid (less than 1%), N-glycoloylneuraminic acid (19%), N-glycoloyl-8-O-methylneuraminic acid (47%), and N-glycoloyl-9-O-acetyl-8-O-methylneuraminic acid (18%). Analysis of sialo-oligomeric material, obtained after mild acid hydrolysis, demonstrated that N-glycoloyl-8-O-methylneuraminic acid can occur as di- and tri-oligomers, linked through the anomeric center and the N-glycoloyl moiety, Neu5Gc8Me-alpha(2----O5)-Neu5Gc8Me and Neu5Gc8Me-alpha(2----O5)-Neu5Gc8Me-alpha (2----O5)-Neu5Gc8Me. Studies on the biosynthesis of N-acyl-8-O-methylneuraminic acid in A rubens, using the tracer S-adenosyl-L-[methyl-14C]methionine, showed that N-acylneuraminate 8-O-methyltransferase activity was present predominantly in the membrane fraction. CMP-N-acetylneuraminic acid monooxygenase activity was found in the soluble protein fraction, in agreement with investigations on the corresponding vertebrate enzyme.
Assuntos
Metiltransferases/metabolismo , Oxigenases de Função Mista/metabolismo , S-Adenosilmetionina/análise , Ácidos Siálicos/biossíntese , Estrelas-do-Mar/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Espectroscopia de Ressonância MagnéticaRESUMO
Lamellar single crystals of amylose V glycerol were grown at 100 degrees C by evaporating water from solutions of amylose in aqueous glycerol. The crystals which were square, with lateral dimensions of several micrometers, gave sharp electron diffraction patterns presenting an orthorhombic symmetry with a probable space group P2(1)2(1)2(1) and unit cell parameters: a = 1.93 +/- 0.01 nm, b = 1.86 +/- 0.01 nm and c (fiber axis) = 0.83 +/- 0.03 nm. The amylose Vglycerol crystal structure which is isomorphous to that of VDMSO consists of an antiparallel pair of left-handed six-fold amylose helices centered on the two-fold screw axes of the cell and probably separated by glycerol molecules. This packing mode is confirmed by de-solvation experiments where the Vglycerol amylose crystals, annealed in ethanol, could be converted into VH amylose without losing their external appearance. The Vglycerol amylose crystals could be seeded by cellulose microfibrils to yield a shish-kebab structure where the amylose crystalline lamellae grew perpendicular to the microfibril directions.
Assuntos
Amilases/química , Cristalografia por Raios X , Glicerol/química , Cristalização , Etanol , Difração de Raios XRESUMO
The influence of several variables on the adsorption of 99mTc(Sn)-diphosphonate complexes on tri-calciumphosphate was determined. The composition of the incubation medium influenced the percentage adsorption: with Hank's balanced salt solution (a medium frequently used for bone cell cultures) and Tris buffer lower percentage adsorption was obtained than with physiological saline as the incubation medium. The influence of addition to the incubation medium of some ions and neutral species, some of which occur in bone fluid, is very specific. Addition of Sn(II) or Mg(II) (a component of HBSS) reduces the amount of adsorption. Addition of Ca(II) and Al(III) had no effect. Addition of sodium-citrate and MDP to the medium and an increase of the pH of the medium decreased the percentage adsorption. The ligand that was used in the preparation of the complex mixture influences the percentage adsorption considerably. The Sn(II) concentration used during the preparation of the 99mTc(Sn)-MDP and 99mTc(Sn)-MHDP complexes showed no definite influence on the percentage adsorption. The pH and ligand concentration, used in the preparation, however, did effect the percentage adsorption. It was concluded that the 99mTc(Sn)-disphosphonate mixtures are part reversibly and part irreversibly bound to tri-calcium-phosphate.
Assuntos
Fosfatos de Cálcio , Ácido Etidrônico , Compostos de Organotecnécio , Medronato de Tecnécio Tc 99m , AdsorçãoRESUMO
The question whether sucrose (Suc) is present inside plastids has been long debated. Low Suc levels were reported to be present inside isolated chloroplasts, but these were argued to be artifacts of the isolation procedures used. We have introduced Suc-metabolizing enzymes in plastids and our experiments suggest substantial Suc entry into plastids. The enzyme levansucrase from Bacillus subtilis efficiently synthesizes fructan from Suc. Targeting of this enzyme to the plastids of tobacco (Nicotiana tabacum) and potato (Solanum tuberosum) plants leads to high-level fructan accumulation in chloroplasts and amyloplasts, respectively. Moreover, introduction of this enzyme in amyloplasts leads to an altered starch structure. Expression of the yeast invertase in potato tuber amyloplasts results in an 80% reduction of total Suc content, showing efficient hydrolysis of Suc by the plastidic invertase. These observations suggest that Suc can enter plastids efficiently and they raise questions as to its function and metabolism in this organelle.