Formation of copper-bleomycin complexes: evidence of a three-step process

The formation of Cu(II)-bleomycin complexes as a function of pH has been studied using circular dichroism, absorption, electron paramagnetic resonance spectroscopy, and potentiometric titration. Our data support the following points: the formation of Cu(II)-bleomycin complexes occurs in a three-step process: a first complex (I) is formed at pH 1.2, which most probably involves the pyrimidine nitrogen, the secondary amine nitrogen, and two water molecules as the four in-plane ligands of copper. A second complex (II) is formed at pH 2.5, through the further coordination of the peptide nitrogen of histidine residue, and histidine imidazole nitrogen giving rise to the release of two protons. The fixation, in apical position, of the alpha-amino nitrogen of beta-aminoalanine occurs in a last step through the release of one additional proton. A value of 2.7 has been obtained for the pK of formation of this third complex, which is the species present at physiological pH. In the Cu(II)-depbleomycin system only one complex (II') has been detected.     

Effects of laser radiation on rhus vernicifera laccase,type 2 Cu-depleted laccase,and stellacyanin

Laser irradiation in the 450 nm region brings about irreversible changes in the copper sites of Rhus vernicifera laccase and its type 2 Cu-depleted derivative. The absorption band at 614 nm disappears after ~ 2 hr of irradiation with a 200 mW laser beam; the amount of the paramagnetic detectable copper does not decrease, indicating no reduction of these types of copper. No apparent rearrangement of the protein backbone occurs, as ultaviolet dichroic spectra of the enzyme before and after the irradiation do not show appreciable differences. Stellacyanin is insensitive to laser radiation at any wavelength.     

Modifications of the molecular weight of membrane-bound nonspecific β-glucosidase in type 1 gaucher disease determined in situ by the radiation inactivation method

The radiation inactivation method has been used to compare the molecular weight of the nonspecific membrane-bound β-glucosidase in situ in normal human spleen and in that of two patients with Gaucher diease type 1. We report, in type 1 Gaucher spleen, the presence of a high molecular weight component (557 000) in addition to the normal low molecular weight component (97 800). The various possible hypotheses explaining this high molecular weight component are discussed.     

Metal anthracycline complexes as a new class of anthracycline derivatives. Pd(II)-adriamycin and Pd(II)-daunorubicin complexes: physicochemical characteristics and antitumor activity

Pd(II) complexes of two anthracyclines, adriamycin and daunorubicin, have been studied. Using potentiometric absorption, fluorescence, and circular dichroism measurements, we have shown that adriamycin can form two complexes with Pd(II). The first complex (I) involves two molecules of drug per Pd(II) ion; one of the molecules is chelated to Pd(II) through the carbonyl oxygen on C12 and the phenolate oxygen on C11, and the other one is bound to Pd(II) through the nitrogen of the amino sugar. This complexation induces a stacking of the two molecules of drug. In the second complex (II), two Pd(II) ions are bound to two molecules of drug (A1 and A2). One Pd(II) is bound to the oxygen on the carbons C11 and C12 of molecule A1 and the amino sugar of molecule A2 whereas the second Pd(II) ion is bound to the oxygen on C11 and C12 of molecule A2 and the amino sugar of molecule A1. The same complexes are formed between Pd(II) and daunorubicin. The stability constant for complex II is beta = (1.3 +/- 0.5) X 10(22). Interaction with DNA has been studied, showing that almost no modification of the complex occurred. This complex displays antitumor activity against P-388 leukemia that compares with that of the free drug. Complex II, unlike adriamycin, does not catalyze the flow of electrons from NADH to molecular oxygen through NADH dehydrogenase.     

Schistosoma mansoni: structural and biochemical characterization of two distinct Venus Kinase Receptors

Venus Kinase Receptors (VKRs) are atypical transmembrane proteins composed of an extracellular Venus FlyTrap module linked through a single helix to a tyrosine kinase domain similar to that of insulin receptors. This structure was first described in Schistosoma mansoni, then in a selected range of invertebrates, including many insects. The preferential expression of VKRs in larvae and gonads suggested their role in development and reproduction. While a single vkr gene was consistently found in all genomes, we identified two distinct vkr genes in S. mansoni. Our data indicated that Smvkr1 and Smvkr2 are very similar in structure and likely originated from gene duplication. Both genes are expressed in all the parasite stages and encode homologous proteins with a conserved VKR structure. Recombinant SmVKR1 and SmVKR2 exhibit tyrosine kinase activities dependent on the binding of distinct small ligand molecules. SmVKR1 and SmVKR2 could represent paralogs with different functions in the parasite.     

Interembryonic regions of the uterus of the viviparous lizard Mabuya brachypoda (Squamata: Scincidae)

Analysis of the structure and physiology of the uterine incubation chambers of viviparous squamates has provided insight concerning adaptations for gestation. However, the literature addressing the biology of the interembryonic regions of the uterus is very limited, presumably because it has been assumed that this area has little role in the development and support of embryos in viviparous squamates. This study was undertaken to examine the histology of the interembryonic regions of Mabuya brachypoda, a viviparous lizard with microlecithal ova and consequently substantial matrotrophic activity. The incubation chambers are oval, distended zones of the uterus, adjacent to the interembryonic regions. The wall of the interembryonic regions includes: mucosa, formed by a cuboidal or columnar epithelium with ciliated and nonciliated cells, and a lamina propria of vascularized connective tissue containing abundant acinar glands; myometrial smooth muscle consisting of inner circular and outer longitudinal layers; and serosa. The segment of the interembryonic region adjacent to the incubation chamber forms a transitional segment that displays folds of the mucosa that protrude into the uterine lumen. The limit of the incubation chamber is well defined by the long mucosal folds of the transitional segment. Long and thin extensions of extraembryonic membranes are present in the lumen of the transitional segment, outside of the incubation chamber region. The presence of abundant uterine glands and extraembryonic membranes in the interembryonic regions during gestation suggests uterine secretory activity and histotrophic transfer of nutrients to embryos in these regions.     

Functional diversity of microboring Ostreobium algae isolated from corals

The filamentous chlorophyte Ostreobium sp. dominates shallow marine carbonate microboring communities, and is one of the major agents of reef bioerosion. While its large genetic diversity has emerged, its physiology remains little known, with unexplored relationship between genotypes and phenotypes (endolithic versus free-living growth forms). Here, we isolated nine strains affiliated to two lineages of Ostreobium (>8% sequence divergence of the plastid gene rbcL), one of which was assigned to the family Odoaceae, from the fast-growing coral host Pocillopora acuta Lamarck 1816. Free-living isolates maintained their bioerosive potential, colonizing pre-bleached coral carbonate skeletons. We compared phenotypes, highlighting shifts in pigment and fatty acid compositions, carbon to nitrogen ratios and stable isotope compositions (δ¹³C and δ¹⁵N). Our data show a pattern of higher chlorophyll b and lower arachidonic acid (20:4ω6) content in endolithic versus free-living Ostreobium. Photosynthetic carbon fixation and nitrate uptake, quantified via 8 h pulse-labeling with ¹³C-bicarbonate and ¹⁵N-nitrate, showed lower isotopic enrichment in endolithic compared to free-living filaments. Our results highlight the functional plasticity of Ostreobium phenotypes. The isotope tracer approach opens the way to further study the biogeochemical cycling and trophic ecology of these cryptic algae at coral holobiont and reef scales.