Deamidation as a widespread phenomenon in two-dimensional polyacrylamide gel electrophoresis of human blood plasma proteins

The human plasma protein patterns obtained by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) is a good model system for post-translational modifications because of the existence of several "ladders" of protein spots [Anderson, N. L., Anderson, N. G., Electrophoresis 1991, 12, 883-906], so-called "trains" of spots. Our investigation of several proteins, amongst others beta2-microglobulin and the haptoglobin chains, found the differences in isoelectric points (p/) to be due to deamidation of asparagines. After enzymatic cleavage with endopeptidases in the 2-D polyacrylamide gel, the asparagine and deamidated asparagine containing peptides were separated and quantified by reversed-phase HPLC. In order to separate these peptides, a neutral pH system was established and, as a result, the differences in hydrophobicity of asparagine-containing and deamidated asparagine-containing peptides increased. But how do deamidated asparagines contribute to the observed spot pattern? One spot in the 2-D gel consists of a mixture of protein species with the same number of deamidated asparagines but on different sequence position sites. The difference between the spots in the "ladder" is a growing number of negative charges introduced in the protein by an increasing number of deamidated asparagines. As a consequence, the mass difference between two spots is exactly 1 Da, which is shown in this paper for intact protein masses and the corresponding deamidated peptides.     

Almost sure Cesàro and Euler summability of sequences of dependent random variables

For a sequence of real random variables C _α-summability is shown under conditions on the variances of weighted sums, comprehending and sharpening strong laws of large numbers (SLLN) of Rademacher-Menchoff and Cramér-Leadbetter, respectively. Further an analogue of Kolmogorov’s criterion for the SLNN is established for E _α-summability under moment and multiplicativity conditions of 4th order, which allows one to weaken Chow’s independence assumption for identically distributed square integrable random variables. The simple tool is a composition of Cesàro-type and of Euler summability methods, respectively. Received: 12 June 2006, Revised: 14 May 2007     

Hydroxylation of sesquiterpenes by enzymes from chicory (Cichorium intybus L.) roots

A microsomal enzyme preparation of chicory roots catalyses the hydroxylation of various sesquiterpene olefins in the presence of NADPH. Most of these hydroxylations take place at an isopropenyl or isopropylidene group. The number of products obtained from any of the substrates is confined to one or, in a few cases, two sesquiterpene alcohols. In addition, the conversion of (+)-valencene into nootkatone through β-nootkatol was observed. The involvement of (+)-germacrene A hydroxylase (a cytochrome P450 enzyme) and other enzymes of sesquiterpene lactone biosynthesis in these reactions is discussed.     

Strigolactones: Plant Hormones with Promising Features

Almost 80 years after the discovery of the first plant hormone, auxin, a few years ago a new class of plant hormones, the strigolactones, was discovered. These molecules have unprecedented biological activity in a number of highly important biological processes in plants but also outside the plant in the rhizosphere, the layer of soil surrounding the roots of plants and teeming with life. The exploitation of this amazing biological activity is not without challenges: the synthesis of strigolactones is complicated and designing the desired activity a difficult task. This minireview describes the current state of knowledge about the strigolactones and how synthetic analogs can be developed that can potentially contribute to the development of a sustainable agriculture.     

Symmetrisierte und zentrierte Folgen von Zufallsvariablen

Ohne Zusammenfassung     

Pressure dependence of the demixing behaviour in polymer blends: I. Experimental procedure

The most common way to influence the liquid-liquid phase behaviour in partially miscible (co-)polymer blends is changing the blending temperature. Since most extruders can handle pressures, up to 300 bar, pressure may also be used to influence the miscibility of polymers during blending. We have developed equipment and an experimental procedure to study the pressure dependence of the liquid-liquid demixing behaviour of high-viscous polymer blends under equilibrium conditions. Small amounts (1–4 grams) of specially made polymers are blended in the ‘DSM MINI EXTRUDER’. After a chosen mixing time, a small portion of the blend is injected into a small capillary tube and kept at the blending temperature. The phase behaviour of the blends as a function of temperature and pressure is studied via laser light scattering (at a scattering angle of 90°) in a specially made 400 bar/250°C window autoclave, where the capillary cell is placed in a high temperature grade silicon oil.     

Assessment of the amount of body water in the Red Knot (Calidris canutus): an evaluation of the principle of isotope dilution with 2H, (17)O, and (18)O as measured with laser spectrometry and isotope ratio mass spectrometry

We have used the isotope dilution technique to study changes in the body composition of a migratory shorebird species (Red Knot, Calidris canutus) through an assessment of the amount of body water in it. Birds were quantitatively injected with a dose of water with elevated concentrations of 2H, (17)O, and (18)O. Thereafter, blood samples were taken and distilled. The resulting water samples were analysed using an isotope ratio mass spectrometry (for 2H and (18)O only) and a stable isotope ratio infrared laser spectrometry (2H, (17)O, and (18)O) to yield estimates of the amount of body water in the birds, which in turn could be correlated to the amount of body fat. Here, we validate laser spectrometry against mass spectrometry and show that all three isotopes may be used for body water determinations. This opens the way to the extension of the doubly labelled water method, used for the determination of energy expenditure, to a triply labelled water method, incorporating an evaporative water loss correction on a subject-by-subject basis or, alternatively, the reduction of the analytical errors by statistically combining the (17)O and (18)O measurements.     

Sugar silanes: versatile reagents for stereocontrolled glycosylation via intramolecular aglycone delivery

A new method for the intramolecular glycosylation of alcohols is described. Utilizing carbohydrate-derived silanes, the catalytic dehydrogenative silylation of alcohols is followed by intramolecular glycosylation. Appropriate combinations of silane position and protecting groups allow highly selective access to β-manno, α-gluco, or β-gluco stereochemical relationships as well as 2-azido-2-deoxy-β-gluco- and 2-deoxy-β-glucosides. Intramolecular aglycone delivery from the C-2 or C-6 position provides 1,2-cis or 1,2-trans glycosides, respectively. Multifunctional acceptor substrates such as hydroxyketones and diols are tolerated and are glycosylated in a site-selective manner.