Pseudobase of malvidin 3-rhamnosido-5-glucoside in am mutants of Pisum sativum

am Mutants of Pisum sativum with white petals have been shown to contain the pigment malvidin 3-rhamnosido-5-glucoside in colourless pseudobase     

Greenish blue flower colour of Strongylodon macrobotrys

The flower colour of Strongyledon macrobotrys is luminous blue green and attracts bats for pollination. The chemical basis for development of the flower colour was investigated. The flower contained an anthocyanin (malvin) and a flavone (saponarin), approximately 1:9 (malvin: saponarin) in molar ratio. The pH of the pigmented epidermal cell sap of the jade vine petal was exceptionally high, 7.90, while the pH value of the colourless inner tissue was 5.60. Copigmentation test using the mixtures of malvin and saponarin (1:9 M ratio) at various pH values revealed that the characteristic blue green colour of the jade vine is developed by copigmentation of malvin with saponarin in slightly alkaline cell sap, pH 7.9. In the copigmentation in slightly alkaline condition, saponarin shows a strong yellow colour, which gives a greenish tone to the flower colour.     

The anthocyanin in blue flowers ofCentaurea cyanus

The anthocyanin in the blue cornflower (Centaurea cyanus) has been known for many years to be cyanidin 3,5-diglucoside, namely cyanin. However, in the course of this study, it became evident that the major anthocyanin in the blue cornflower is not cyanin but cyanidin 3-succinyl glucoside 5-glucoside. This anthocyanin has not been reported in the literature and is tentatively called “centaurocyanin”. Centaurocyanin is chromatographically identical with the anthocyanin contained in crystalline protocyanin, the blue pigment from the cornflower. thus, there seems no doubt that this anthocyanin, but not cyanin, forms the blue complex pigment protocyanin.     

Anthocyanins of some Malaysian members of the gesneriaceae

The occurrence of 'normal' 3-hydroxylated anthocyanins in 8 Malaysian species of the Gesneriaceae supports the important chemotaxonomic results for this family. New compounds found in Chirita, Didissandra and Didymocarpus are the 3-arabinosylglucoside-5-glucosides of cyanidin and malidin, pigments which may have some systematic value.     

Participation of p-coumaroyl glucose residue of awobanin in synthesis of the complex molecule of commelinin

In Vitro synthesis of commelinin-like metallo-anthocyanins was attempted using flavocommelin, metals and some anthocyanins having structures similar to awobanin, i.e. delphin, violanin, shisonin and cyanin. Only shisonin formed stable metallo-anthocyanins (with Mg, Zn, Cd, Mn, Ni and Co); the blue pigments were homologues of those prepared from awobanin. The ineffectiveness of delphin (deacylated awobanin) and cyanin (deacylated shisonin) suggests that the p-coumaroyl glucose residues of awobanin and of shisonin play an important role in the formation of rhamnosylglucoside) indicates that the length and nature of the sugar chain are critical factors for the formation of The ineffectiveness of violanin, similar to awobanin except for a different p-coumaroyl sugar chain (p-coumaroyl rhamnosylglucoside) indicates that the length and nature of the sugar chain are critical factors for the formation of commelinin and similar complex pigments.     

Covalent anthocyanin-flavone dimer from leaves of Oxalis triangularis

The anthocyanin-flavone C-glycoside, (malvidin 3-O-(6(II)-O-alpha-rhamnopyranosyl(AIV)-beta-glucopyranoside(AII))-5-O-beta-glucopyranoside(AIII)) (apigenin 6-C-(2(II)-O-beta-glucopyranosyl(FIII)-beta-glucopyranoside(FII))) malonate(AV) (A(IV)-4-->A(V)-1, F(III)-6-->A(V)-3) (1), has been isolated from leaves of Oxalis triangularis A. St.-Hil. In the 1D (1)H NMR spectrum of 1 dissolved in CD(3)OD-CF(3)CO(2)D (95:5), MTFA, recorded 45 min after sample preparation, this covalently linked dimer occurred mainly as flavylium cation (38%) and two equilibrium forms assigned to be quinonoidal bases (54%), whereas only minor amounts of the hemiacetal forms were present. After five days storage at 300 K, the hemiacetals (39%) and flavylium cation (38%) constituted the main forms of 1. More simple anthocyanins are normally considered to be on the flavylium cation form in acidified deuterated methanol. The cross-peaks observed in NOESY NMR spectra of 1 indicated the presence of vertical 'pi-pi' stacking between the B-ring of the flavone unit and the A-ring of each of the two forms assigned to be quinonoidal bases. It was not possible to discriminate between inter- or intramolecular association mechanisms. The equilibria between the various forms of 1 were studied by two-dimensional NOESY and ROESY NMR spectroscopy. 2D HSQC-TOCSY NMR spectroscopy was among the methods used for characterization of the various forms.     

Morphological responses to high sugar concentrations differ from adaptation to high salt concentrations in the xerophilic fungi Wallemia spp.

Fungi from the food-borne basidiomycetous genus Wallemia, which comprises Wallemia ichthyophaga, Wallemia muriae and Wallemia sebi, are among the most xerophilic organisms described. Their morphological adaptations to life at high NaCl concentrations are reflected in increased cell-wall thickness and size of cellular aggregates. The objectives of this study were to examine their growth and to define cell morphology and any ultrastructural cell-wall changes when these fungi are grown in low and high glucose and honey concentrations, as environmental osmolytes. We analysed their growth parameters and morphological characteristics by light microscopy and transmission and scanning electron microscopy.Wallemia ichthyophaga grew slowly in all of the sugar-based media, while W. muriae and W. sebi demonstrated better growth. Wallemia ichthyophaga adapted to the high glucose and honey concentrations with formation of larger cellular aggregates, while cell-wall thickness was increased only at the high glucose concentration. Wallemia muriae and W. sebi demonstrated particularly smaller sizes of hyphal aggregates at the high glucose concentration, and different and less explicit changes in cell-wall thickness. Adaptive responses show that the phylogenetically more distant W. ichthyophaga is better adapted to high salt conditions, whereas W. muriae and W. sebi cope better with a high sugar environment.     

Sphagnum re-introduction in degraded peatlands: The effects of aggregation,species identity and water table

In European peatlands which have been drained and cut-over in the past, re-vegetation often stagnates after the return of a species-poor Sphagnum community. Re-introduction of currently absent species may be a useful tool to restore a typical, and more diverse, Sphagnum vegetation and may ultimately improve the functioning of peatland ecosystems, regarding atmospheric carbon sequestration. Yet, the factors controlling the success of re-introduction are unclear. In Ireland and Estonia, we transplanted small and large aggregates of three Sphagnum species into existing vegetation. We recorded changes in cover over a 3-year period, at two water levels (?5 and ?20 cm).Performance of transplanted aggregates of Sphagnum was highly species specific. Hummock species profited at low water tables, whereas hollow species profited at high water tables. But our results indicate that performance and establishment of species was also promoted by increased aggregate size. This mechanism (positive self-association) has earlier been seen in other ecosystems, but our results are the first to show this mechanism in peatlands. Our results do not agree with present management, which is aimed at retaining water on the surface of peat remnants in order to restore a functional and diverse Sphagnum community. More than the water table, aggregate size of the reintroduced species is crucial for species performance, and ultimately for successful peatland restoration.     

Investigation of the in vivo organization of anthocyanins using resonance raman microspectrometry

Resonance Raman (RR) microspectrometry constitutes a new means for studying the organization of anthocyanins in living tissues. RR spectra of the pigments present in a single cell have been recorded from the skins of the mature berries of the ‘Pinot noir’ grape, as well as the petals of the common mallow. Comparison of these spectra with those obtained from model solutions of anthocyanins permits us to conclude that in the ‘Pinot noir’ berries, malvidin 3-glucoside is the main pigment. Furthermore, inside the skin it is essentially in the quinonoidal base form, whereas in the outer face of the skin it is mainly in the flavylium form. In the upper epidermis of petals of the common mallow, only malvidin 3,5-diglucoside could be detected, entirely in the cationic flavylium form. Since self-association or co-pigmentation processes do not seem to contribute much to the RR spectra, we conclude that they perturb the electronic excited state of the monomeric anthocyanin chromophores much more than they modify the corresponding electronic ground state. In the case of the skins of the mature berries of the ‘Pinot noir’ grape, dark grains, which we believe to be anthocyanoplasts, have been observed. In vivo RR spectra of the dark grains have been recorded.     

The temperature-dependent self-association of beta-lactoglobulin C in glycine buffers

The self-association of β-lactoglobulin C at low pH (ca. 2.5) in glycine buffers has been studied at four temperatures, 10, 16, 20, and 25 °C, by low- and high-speed sedimentation equilibrium experiments. One buffer had an ionic strength of 0.1 and the other an ionic strength of 0.2. With either buffer the concentration dependence of the apparent weight average molecular weight, M_wa, was characteristic of a nonideal self-association. Like its genetic variants, β-lactoglobulin A and B, the self-association of β-lactoglobulin C increased with decreasing temperature; however, at the same temperature the association was always stronger in the buffer having the higher ionic strength. Several models were used to test the self-association, and a monomer-dimer self-association seemed to describe the self-association best with either buffer. Values of the association equilibrium constant, K₂, and the second virial coefficient, BM₁, are reported at each temperature for both series of experiments. Values of the thermodynamic functions, ΔG °, ΔH °, and ΔS °, are also reported for these experiments.     

The structure of a polymer containing galactosamine from walls of Bacillus subtilis 168. (Short Communication)

Cell walls of Bacillus subtilis 168 contain a phosphorylated polysaccharide composed of N-acetylgalactosamine, glucose and phosphate. Controlled acid hydrolysis gives 3-O-beta-d-glucopyranosyl-N-acetylgalactosamine with a phosphomonoester group at the 6-position of glucose. It is likely that the polysaccharide consists of disaccharide units connected by phosphodiester residues joining the 1-position of N-acetylgalactosamine to the 6-position of glucose in the neighbouring unit.     

Co-pigmentation and the color change with age in petals ofFuchsia hybrida

Pure or genuine malvin was isolated fromFuchsia petals for the first time and characterized as the malvin anhydro-base. The conditions for the co-pigmentation were examinedin vitro with regard to anthocyanin, co-pigment and pH, and the co-pigmentation occurred as the result of interaction between anthocyanin and co-pigment without any participation of metallic elements. The blue-violet color of youngFuchsia petals appeared at pH 4.8 in the 1∶0.6 molar ratio of anthocyanin to co-pigments. The color change from blue-violet in young petals to purple-red in old ones was caused by co-pigmentation and the pH change from 4.8 to 4.2. The decrease of pH in the old petals was due to the increase of organic acids such as aspartic, malic and tartaric acids.     

Weak self-association of human growth hormone investigated by nitrogen-15 NMR relaxation

The self-association of human growth hormone(hGH) was investigated using 15N NMR relaxation.The investigation relies on the 15N R1 and R2 relaxation rates and the heteronuclear{1H}-15N NOEs of the backbone amide groups at multiple protein concentrations. It is shown that the rotational correlation time of hGH in solution depends strongly on its concentration, indicating a significant degree of self-association.The self-association is reversible and the monomers in the aggregates are noncovalently linked. Extrapolation of the relaxation data to zero concentration predicts a correlation time of 13.4 ns and a rotational diffusion anisotropy of 1.26 for monomeric hGH, in agreement with the rotational diffusion properties estimated by hydrodynamic calculations. Moreover, the extrapolation allows characterization of the backbone dynamics of monomeric hGH without interference from self-association phenomena, and it is found that hGH is considerably more flexible than originally thought. A concerted least-squares analysis of the 15N relaxations and their concentration dependence reveals that the self-association goes beyond a simple monomer-dimer equilibrium, and that tetramers or other multimeric states co-exist in fast exchange with the monomeric and dimeric hGH at sub-millimolar concentrations. Small changes in the 1H and 15N amide chemical shifts suggest that a region around the C-terminus is involved in the oligomer formation.     

On the interaction of caffeine with nucleic acids: III. 1NMR studies of caffeine-5'-adenosine monophosphate and caffeine-poly(riboadenylate) interactions

1) The self-association of both caffeine (Cf) and 5'-adenosine monophosphate (AMP) in aqueous solution has been reinvestigated by ¹H NMR. The self-association process is characterized by an isodesmic model. The apparent self-association constants of the vertical stacking process are K_Cf= (10.6 ± 1.0) M^?1 and k_amp = (1.67 ± 0.17) M^?1. The arrangement of the monomeric units in the stacked aggregates is discussed in terms of isoshielding curves theoretically calculated by Giessner-Prettre and Pullman. Models are proposed which are consistent with these and further previous NMR data. 2) The interaction of Cf and AMP has been studied by ^?1 H NMR. The apparent association constant of the complex Cf-AMP is K_C-A = (7.3 ± 1.2) M^?1.Two models of the mutual arrangement of AMP and Cf in the complex are proposed on the basis of the calculated isoshielding curves considering both ring current and local atomic diamagnetic anisotropy effects. 3) The interaction of Cf and poly(riboadenylate), (rA)_n is indicated by a downfield shift of the H-8 line but an upfield shift of the H-2 line in the ¹H NMR spectra of (rA)_n. The concentration dependence of the ¹H NMR shifts of both Cf and (rA)_n can be explained by the existence of two binding mechanisms. We suggest (i) partial insertion of Cf between adjacent base residues of ordered single-stranded regions of (rA)_n and (ii) outside binding of Cf in form of monomeric Cf as well as of self-associated aggregates. The complex geometry of insertion proposed on the basis of the calculated isoshielding curves is characterized by a stronger overlapping of the Cf ring and the H-2 proton of (rA)_n as compared to the H-8 proton.     

Composition of gum exudates from Anacardium occidentale

The composition and solution properties of Indian and Papuan specimens of the gum from Anacardium occidentale have been studied and found to be closely similar. Contrary to earlier reports by Indian workers, this gum does not contain galacturonic acid. It does, however, contain glucose; this appears to be the first report of the presence of this sugar in a plant gum exudate. A freeze-dried sample of the gum was examined at intervals over a period of 2 months; its weight-average MW increased by a factor of three in that time, and molecular-sieve chromatography showed that self-association occurred with the formation of a small proportion of a very high MW component.     

Discovery of N-methyl-4-(4-methoxyanilino)quinazolines as potent apoptosis inducers. Structure–activity relationship of the quinazoline ring

As a continuation of our efforts to discover and develop apoptosis inducing N-methyl-4-(4-methoxyanilino)quinazolines as novel anticancer agents, we explored substitution at the 5-, 6-, 7-positions of the quinazoline and replacement of the quinazoline by other nitrogen-containing heterocycles. A small group at the 5-position was found to be well tolerated. At the 6-position a small group like an amino was preferred. Substitution at the 7-position was tolerated much less than at the 6-position. Replacing the carbon at the 8-position or both the 5- and 8-positions with nitrogen led to about 10-fold reductions in potency. Replacement of the quinazoline ring with a quinoline, a benzo[d][1,2,3]triazine, or an isoquinoline ring showed that the nitrogen at the 1-position is important for activity, while the carbon at the 2-position can be replaced by a nitrogen and the nitrogen at the 3-position can be replaced by a carbon. Through the SAR study, several 5- or 6-substituted analogs, such as 2a and 2c, were found to have potencies approaching that of lead compound N-(4-methoxyphenyl)-N,2-dimethylquinazolin-4-amine (1g, EP128495, MPC-6827, Azixa®).     

Thermodynamical model of indefinite mixed association of two components and NMR data analysis for caffeine-AMP interaction

This paper describes the model used to estimate the parameters of caffeine-AMP interactions from corresponding 1H-NMR measurements and some methods of data analysis by which the applicability of the model has been checked. The model of mixed association is applicable to a mixture of any two substances A and C which exhibit indefinite aggregates in both self-association and mixed association. In aggregates, only nearest neighbour interaction is assumed. The model is described by three equilibrium constants: Kaa and Kcc (for self-association of A, or C, respectively), and Kac (for mixed association).     

Ribbon shaped histone H4 aggregates

The self-association process of histone H4 molecules into large aggregates is highly cooperative at an ionic strength of 0.03. Carefully prepared aggregates at this ionic strength are ribbon-shaped and take the form of right-handed helices. These ribbons have a uniform width of 20.5nm. The overall diameter of the “cylinder” formed by the helix is ca. 19.5nm, the average pitch of the helices 20nm, and the mean contour length 720nm. The thickness of the ribbon is less than 3nm.     

Bruton’s tyrosine kinase phosphorylates scaffolding and RNA-binding protein G3BP1 to induce stress granule aggregation during host sensing of foreign ribonucleic acids

The Ras-GTPase activating protein SH3 domain-binding protein 1 (G3BP1) plays a critical role in the formation of classical and antiviral stress granules in stressed and virus-infected eukaryotic cells, respectively. While G3BP1 is known to be phosphorylated at serine residues which could affect stress granule assembly, whether G3BP1 is phosphorylated at tyrosine residues and how this posttranslational modification might affect its functions is less clear. Here, we show using immunoprecipitation and immunoblotting studies with 4G10 antibody that G3BP1 is tyrosine-phosphorylated when cells are stimulated with the synthetic double-stranded RNA analog polyinosinic:polycytidylic acid to mimic viral infection. We further demonstrate via co-immunoprecipitation and inhibitor studies that Bruton’s tyrosine kinase (BTK) binds and phosphorylates G3BP1. The nuclear transport factor 2–like domain of G3BP1 was previously shown to be critical for its self-association to form stress granules. Our mass spectrometry, mutational and biochemical cross-linking analyses indicate that the tyrosine-40 residue in this domain is phosphorylated by BTK and critical for G3BP1 oligomerization. Furthermore, as visualized via confocal microscopy, pretreatment of cells with the BTK inhibitor LFM-A13 or genetic deletion of the btk gene or mutation of G3BP1-Y40 residue to alanine or phenylalanine all significantly attenuated the formation of antiviral stress granule aggregates upon polyinosinic:polycytidylic acid treatment. Taken together, our data indicate that BTK phosphorylation of G3BP1 induces G3BP1 oligomerization and facilitates the condensation of ribonucleoprotein complexes into macromolecular aggregates.