Binding of Antitumor Ruthenium(III) Complexes to Plasma Proteins

Presently, there is large interest in analysing the interactions in vitro with plasma proteins of some novel antitumor ruthenium(III) complexes that are in preclinical or clinical phase. The joint application of separation and spectroscopic techniques provides valuable information on the nature and the properties of the resulting ruthenium/protein adducts. Recent work carried out in our laboratory points out that, under physiological conditions, some selected ruthenium(III) complexes bind plasma proteins tightly with a marked preference for surface imidazole groups. Representative examples of interactions of antitumor ruthenium(III) complexes with plasma proteins such as albumin and transferrin are given. Notably the antitumor ruthenium(III) complexes considered here bind proteins much tighter than DNA; it is proposed that protein binding of ruthenium(III) complexes will have a large impact on the biodistribution, the pharmacokinetics and the mechanism of action of these experimental drugs

New insight into the atmospheric chloromethane budget gained using gained using

International audienceAtmospheric chloromethane (CH3Cl) plays an important role in stratospheric ozone destruction, but many uncertainties still exist regarding strengths of both sources and sinks and the processes leading to formation of this naturally occurring gas. Recent work has identified a novel chemical origin for CH3Cl, which can explain its production in a variety of terrestrial environments: The widespread structural component of plants, pectin, reacts readily with chloride ion to form CH3Cl at both ambient and elevated temperatures (Hamilton et al., 2003). It has been proposed that this abiotic chloride methylation process in terrestrial environments could be responsible for formation of a large proportion of atmospheric CH3Cl. However, more information is required to determine the global importance of this new source and its contribution to the atmospheric CH3Cl budget. A potentially powerful tool in studying the atmospheric CH3Cl budget is the use of stable carbon isotope ratios. In an accompanying paper it is reported that the reaction of CH3Cl with OH radical, the dominant sink for atmospheric CH3Cl, is accompanied by an unexpectedly large fractionation factor (Gola et al., 2005). Another recently published study shows that CH3Cl formed by the abiotic methylation process at ambient temperatures has a unique stable carbon isotope signature, extremely depleted in 13C, unequivocally distinguishing it from all other known sources (Keppler et al., 2004). Using these findings together with data existing in the literature, we here present three scenarios for an isotopic mass balance for atmospheric CH3Cl. Our calculations provide strong support for the proposal that the bulk fraction of atmospheric CH3Cl (1.8 to 2.5Tg yr?1) is produced by an abiotic chloride methylation process in terrestrial ecosystems, primarily located in tropical and subtropical areas, where turnover of biomass is highest. Furthermore our calculations also indicate that the microbial soil sink for CH3Cl is likely to be much larger (>1Tg yr?1) than that previously assumed

New insight into the atmospheric chloromethane budget gained using stable carbon isotope ratios

International audienceAtmospheric chloromethane (CH3Cl) plays an important role in stratospheric ozone destruction, but many uncertainties still exist regarding strengths of both sources and sinks and the processes leading to formation of this naturally occurring gas. Recent work has identified a novel chemical origin for CH3Cl, which can explain its production in a variety of terrestrial environments: the widespread structural component of plants, pectin, reacts readily with chloride ion to form CH3Cl at both ambient and elevated temperatures (Hamilton et al., 2003). It has been proposed that this abiotic chloride methylation process in terrestrial environments could be responsible for formation of a large proportion of atmospheric CH3Cl. However, more information is required to determine the global importance of this new source and its contribution to the atmospheric CH3Cl budget. A potentially powerful tool in studying the atmospheric CH3Cl budget is the use of stable carbon isotope ratios. In an accompanying paper it is reported that the reaction of CH3Cl with OH radical, the dominant sink for atmospheric CH3Cl, is accompanied by an unexpectedly large fractionation factor (Gola et al., 2005). Another recently published study shows that CH3Cl formed by the abiotic methylation process at ambient temperatures has a unique stable carbon isotope signature, extremely depleted in 13C, unequivocally distinguishing it from all other known sources (Keppler et al., 2004). Using these findings together with data existing in the literature, we here present three scenarios for an isotopic mass balance for atmospheric CH3Cl. Our calculations provide strong support for the proposal that the largest source of atmospheric CH3Cl (1800 to 2500 Gg yr-1) is the abiotic methylation of chloride in terrestrial ecosytems, primarily located in tropical and subtropical areas where turnover of biomass is highest. Furthermore our calculations also indicate that the microbial soil sink for CH3Cl is likely to be much larger (>1000 Gg yr-1) than that previously assumed

Comparison of the Antiproliferative Activity of Two Antitumour Ruthenium(III) Complexes With Their Apotransferrin and Transferrin-Bound Forms in a Human Colon Cancer Cell Line

Two ruthenium(III) complexes, namely trans-indazolium[tetrachlorobis(indazole)- ruthenate(III)], HInd[RuInd2Cl4] and trans-imidazolium[tetrachlorobis(imidazole)- ruthenate(III)], HIm[RuIm2Cl4] exhibit high anticancer activity in an autochthonous colorectal carcinoma model in rats. Recently, it has been shown that both complexes bind specifically to human serum apotransferrin and the resulting adducts have been studied through spectroscopic and chromatographic techniques with the ultimate goal of preparing adducts with good selectivity for cancer cells due to the fact that tumour cells express high amounts of transferrin receptors on their cell surface

The Biochemical Role of the Heat Shock Protein 90 Chaperone Complex in Establishing Human Telomerase Activity

Telomerase is a ribonucleoprotein complex that synthesizes the G-rich DNA found at the 3'-ends of linear chromosomes. Human telomerase consists minimally of a catalytic protein (hTERT) and a template-containing RNA (hTR), although other proteins are involved in regulating telomerase activity in vivo. Several chaperone proteins, including hsp90 and p23, have demonstrable roles in establishing telomerase activity both in vitro and in vivo, and previous reports indicate that hsp90 and p23 are required for the reconstitution of telomerase activity from recombinant hTERT and hTR. Here we report that hTERT and hTR associate in the absence of a functional hsp90-p23 heterocomplex. We also report that hsp90 inhibitors geldanamycin and novobiocin inhibit recombinant telomerase even after telomerase is assembled. Inhibition by geldanamycin could be overcome by allowing telomerase to first bind its primer, suggesting a role for hsp90 in loading telomerase onto the telomere. Inhibition by novobiocin could not similarly be overcome but instead resulted in destabilization of the hTERT polypeptide. We propose that the hsp90-p23 complex fine tunes and stabilizes a functional telomerase structure, allowing primer loading and extension

New insight into the atmospheric chloromethane budget gained using stable carbon isotope ratios

Atmospheric chloromethane (CH3Cl) plays an important role in stratospheric ozone destruction, but many uncertainties still exist regarding strengths of both sources and sinks and the processes leading to formation of this naturally occurring gas. Recent work has identified a novel chemical origin for CH3Cl, which can explain its production in a variety of terrestrial environments: the widespread structural component of plants, pectin, reacts readily with chloride ion to form CH3Cl at both ambient and elevated temperatures (Hamilton et al., 2003). It has been proposed that this abiotic chloride methylation process in terrestrial environments could be responsible for formation of a large proportion of atmospheric CH3Cl. However, more information is required to determine the global importance of this new source and its contribution to the atmospheric CH3Cl budget. A potentially powerful tool in studying the atmospheric CH3Cl budget is the use of stable carbon isotope ratios. In an accompanying paper it is reported that the reaction of CH3Cl with OH radical, the dominant sink for atmospheric CH3Cl, is accompanied by an unexpectedly large fractionation factor (Gola et al., 2005). Another recently published study shows that CH3Cl formed by the abiotic methylation process at ambient temperatures has a unique stable carbon isotope signature, extremely depleted in 13C, unequivocally distinguishing it from all other known sources (Keppler et al., 2004). Using these findings together with data existing in the literature, we here present three scenarios for an isotopic mass balance for atmospheric CH3Cl. Our calculations provide strong support for the proposal that the largest source of atmospheric CH3Cl (1800 to 2500 Gg yr-1) is the abiotic methylation of chloride in terrestrial ecosytems, primarily located in tropical and subtropical areas where turnover of biomass is highest. Furthermore our calculations also indicate that the microbial soil sink for CH3Cl is likely to be much larger (>1000 Gg yr-1) than that previously assumed

ortho-Quinone tanshinones directly inhibit telomerase through an oxidative mechanism mediated by hydrogen peroxide

The tanshinone natural products possess a variety of pharmacological properties including anti-bacterial, anti-inflammatory, anti-oxidant, and anti-neoplastic activity. The molecular basis of these effects, however, remains largely unknown. In the present study, we explored the direct effect of tanshinones on the enzyme telomerase. Telomerase is up-regulated in the majority of cancer cells and is essential for their survival, making it a potential anti-cancer drug target. We found that the ortho-quinone tanshinone II-A inhibits telomerase in a time- and DTT-dependent fashion, and the hydrogen peroxide scavenger catalase protected telomerase from inactivation. These findings demonstrate that ortho-quinone containing tanshinones can inhibit telomerase owing to their ability to generate reactive oxygen species. The results also provide evidence that telomerase is directly and negatively regulated by reactive oxygen species

Controlled assembly of SNAP-PNA-fluorophore systems on DNA templates to produce fluorescence resonance energy transfer

The SNAP protein is a widely used self-labeling tag that can be used for tracking protein localization and trafficking in living systems. A model system providing controlled alignment of SNAP-tag units can provide a new way to study clustering of fusion proteins. In this work, fluorescent SNAP-PNA conjugates were controllably assembled on DNA frameworks forming dimers, trimers, and tetramers. Modification of peptide nucleic acid (PNA) with the O6-benzyl guanine (BG) group allowed the generation of site-selective covalent links between PNA and the SNAP protein. The modified BG-PNAs were labeled with fluorescent Atto dyes and subsequently chemo-selectively conjugated to SNAP protein. Efficient assembly into dimer and oligomer forms was verified via size exclusion chromatography (SEC), electrophoresis (SDS-PAGE), and fluorescence spectroscopy. DNA directed assembly of homo- and hetero-dimers of SNAP-PNA constructs induced homo- and hetero-FRET, respectively. Longer DNA scaffolds controllably aligned similar fluorescent SNAP-PNA constructs into higher oligomers exhibiting homo-FRET. The combined SEC and homo-FRET studies indicated the 1:1 and saturated assemblies of SNAP-PNA-fluorophore:DNA formed preferentially in this system. This suggested a kinetic/stoichiometric model of assembly rather than binomially distributed products. These BG-PNA-fluorophore building blocks allow facile introduction of fluorophores and/or assembly directing moieties onto any protein containing SNAP. Template directed assembly of PNA modified SNAP proteins may be used to investigate clustering behavior both with and without fluorescent labels which may find use in the study of assembly processes in cells

A novel EGFR inhibitor acts as potent tool for hypoxia-activated prodrug systems and exerts strong synergistic activity with VEGFR inhibition in vitro and in vivo

Small-molecule EGFR inhibitors have distinctly improved the overall survival especially in EGFR-mutated lung cancer. However, their use is often limited by severe adverse effects and rapid resistance development. To overcome these limitations, a hypoxia-activatable Co(III)-based prodrug (KP2334) was recently synthesized releasing the new EGFR inhibitor KP2187 in a highly tumor-specific manner only in hypoxic areas of the tumor. However, the chemical modifications in KP2187 necessary for cobalt chelation could potentially interfere with its EGFR-binding ability. Consequently, in this study, the biological activity and EGFR inhibition potential of KP2187 was compared to clinically approved EGFR inhibitors. In general, the activity as well as EGFR binding (shown in docking studies) was very similar to erlotinib and gefitinib (while other EGFR-inhibitory drugs behaved different) indicating no interference of the chelating moiety with the EGFR binding. Moreover, KP2187 significantly inhibited cancer cell proliferation as well as EGFR pathway activation in vitro and in vivo. Finally, KP2187 proved to be highly synergistic with VEGFR inhibitors such as sunitinib. This indicates that KP2187releasing hypoxia-activated prodrug systems are promising candidates to overcome the clinically observed enhanced toxicity of EGFR-VEGFR inhibitor combination therapies

C–O–H–S fluids and granitic magma : how S partitions and modifies CO2 concentrations of fluid-saturated felsic melt at 200 MPa

Author Posting. © The Author(s), 2011. This is the author's version of the work. It is posted here by permission of Springer for personal use, not for redistribution. The definitive version was published in Contributions to Mineralogy and Petrology 162 (2011): 849-865, doi:10.1007/s00410-011-0628-1.Hydrothermal volatile-solubility and partitioning experiments were conducted with fluid-saturated haplogranitic melt, H2O, CO2, and S in an internally heated pressure vessel at 900°C and 200 MPa; three additional experiments were conducted with iron-bearing melt. The run-product glasses were analyzed by electron microprobe, FTIR, and SIMS; and they contain ≤ 0.12 wt% S, ≤ 0.097 wt.% CO2, and ≤ 6.4 wt.% H2O. Apparent values of log ƒO2 for the experiments at run conditions were computed from the [(S6+)/(S6++S2-)] ratio of the glasses, and they range from NNO-0.4 to NNO+1.4. The C-O-H-S fluid compositions at run conditions were computed by mass balance, and they contained 22-99 mol% H2O, 0-78 mol% CO2, 0-12 mol% S, and < 3 wt% alkalis. Eight S-free experiments were conducted to determine the H2O and CO2 concentrations of melt and fluid compositions and to compare them with prior experimental results for C-O-H fluid-saturated rhyolite melt, and the agreement is excellent. Sulfur partitions very strongly in favor of fluid in all experiments, and the presence of S modifies the fluid compositions, and hence, the CO2 solubilities in coexisting felsic melt. The square of the mole fraction of H2O in melt increases in a linear fashion, from 0.05-0.25, with the H2O concentration of the fluid. The mole fraction of CO2 in melt increases linearly, from 0.0003-0.0045, with the CO2 concentration of C-O-H-S fluids. Interestingly, the CO2 concentration in melts, involving relatively reduced runs (log ƒO2 ≤ NNO+0.3) that contain 2.5-7 mol% S in the fluid, decreases significantly with increasing S in the system. This response to the changing fluid composition causes the H2O and CO2 solubility curve for C-O-H-S fluid-saturated haplogranitic melts at 200 MPa to shift to values near that modeled for C-O-H fluid-saturated, S-free rhyolite melt at 150 MPa. The concentration of S in haplogranitic melt increases in a linear fashion with increasing S in C-O-H-S fluids, but these data show significant dispersion that likely reflects the strong influence of ƒO2 on S speciation in melt and fluid. Importantly, the partitioning of S between fluid and melt does not vary with the (H2O/H2O+CO2) ratio of the fluid. The fluid-melt partition coefficients for H2O, CO2, and S and the atomic (C/S) ratios of the run-product fluids are virtually identical to thermodynamic constraints on volatile partitioning and the H, S, and C contents of pre-eruptive magmatic fluids and volcanic gases for subduction-related magmatic systems thus confirming our experiments are relevant to natural eruptive systems.This research was supported in part by National Science Foundation awards EAR 0308866 and EAR-0836741 to J.D.W