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      Copper(II) Complexes with Mixed Heterocycle Ligands as Promising Antibacterial and Antitumor Species

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          Abstract

          Complexes with mixed ligands [Cu(N-N) 2(pmtp)](ClO 4) 2 (( 1) N-N: 2,2′-bipyridine; ( 2) L: 1,10-phenanthroline and pmpt: 5-phenyl-7-methyl-1,2,4-triazolo[1,5- a]pyrimidine) were synthesized and structurally and biologically characterized. Compound ( 1) crystallizes into space group Pa and ( 2) in P-1. Both complexes display an intermediate stereochemistry between the two five-coordinated ones. The biological tests indicated that the two compounds exhibited superoxide scavenging capacity, intercalative DNA properties, and metallonuclease activity. Tests on various cell systems indicated that the two complexes neither interfere with the proliferation of Saccharomyces cerevisiae or BJ healthy skin cells, nor cause hemolysis in the active concentration range. Nevertheless, the compounds showed antibacterial potential, with complex ( 2) being significantly more active than complex ( 1) against all tested bacterial strains, both in planktonic and biofilm growth state. Both complexes exhibited a very good activity against B16 melanoma cells, with a higher specificity being displayed by compound ( 1). Taken together, the results indicate that complexes ( 1) and ( 2) have specific biological relevance, with potential for the development of antitumor or antimicrobial drugs.

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          Mixed-ligand complexes of ruthenium(II): factors governing binding to DNA

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            New and Redesigned pRS Plasmid Shuttle Vectors for Genetic Manipulation of Saccharomyces cerevisiae

            We have constructed a set of 42 plasmid shuttle vectors based on the widely used pRS series for use in the budding yeast Saccharomyces cerevisiae and the bacterium Escherichia coli. This set of pRSII plasmids includes new shuttle vectors that can be used with histidine and adenine auxotrophic laboratory yeast strains carrying mutations in the genes HIS2 and ADE1 , respectively. Our pRSII plasmids also include updated versions of commonly used pRS plasmids from which common restriction sites that occur within their yeast-selectable biosynthetic marker genes have been removed to increase the availability of unique restriction sites within their polylinker regions. Hence, our pRSII plasmids are a complete set of integrating, centromere and 2μ episomal plasmids with the biosynthetic marker genes ADE2 , HIS3 , TRP1 , LEU2 , URA3 , HIS2 , and ADE1 and a standardized selection of at least 16 unique restriction sites in their polylinkers. Additionally, we have expanded the range of drug selection options that can be used for PCR-mediated homologous replacement using pRS plasmid templates by replacing the G418-resistance kanMX4 cassette of pRS400 with MX4 cassettes encoding resistance to phleomycin, hygromycin B, nourseothricin, and bialaphos. Finally, in the process of generating the new plasmids, we have determined several errors in existing publicly available sequences for several commonly used yeast plasmids. Using our updated sequences, we constructed pRS plasmid backbones with a unique restriction site for inserting new markers to facilitate future expansion of the pRS series.
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              Nuclease activity gives an edge to host-defense peptide piscidin 3 over piscidin 1, rendering it more effective against persisters and biofilms

              Host-defense peptides (HDPs) feature evolution-tested potency against life-threatening pathogens. While piscidin 1 (p1) and piscidin 3 (p3) are homologous and potent fish HDPs, only p1 is strongly membranolytic. Here, we hypothesize that another mechanism imparts p3 strong potency. We demonstrate that the N-termini of both peptides coordinate Cu 2+ and p3-Cu cleaves isolated DNA at a rate on par with free Cu 2+ but significantly faster than p1-Cu. On planktonic bacteria, p1 is more antimicrobial but only p3 features copper-dependent DNA cleavage. On biofilms and persister cells, p3-Cu is more active than p1-Cu, commensurate with stronger peptide-induced DNA damage. Molecular dynamics and NMR show that more DNA-peptide interactions exist with p3 than p1, and the peptides adopt conformations simultaneously poised for metal- and DNA-binding. These results generate several important conclusions. First, homologous HDPs cannot be assumed to have identical mechanisms since p1 and p3 eradicate bacteria through distinct relative contributions of membrane and DNA-disruptive effects. Second, the nuclease and membrane activities of p1 and p3 show that naturally occurring HDPs can inflict not only physicochemical but also covalent damage. Third, strong nuclease activity is essential for biofilm and persister cell eradication, as shown by p3, the homolog more specific toward bacteria and more expressed in vascularized tissues. Fourth, p3 combines several physicochemical properties (e.g., Amino Terminal Copper and Nickel binding motif; numerous arginines; moderate hydrophobicity) that confer low membranolytic effects, robust copper-scavenging capability, strong interactions with DNA, and fast nuclease activity. This new knowledge could help design novel therapeutics active against hard-to-treat persister cells and biofilms.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Molecules
                Molecules
                molecules
                Molecules
                MDPI
                1420-3049
                19 August 2020
                September 2020
                : 25
                : 17
                : 3777
                Affiliations
                [1 ]National Institute of Materials Physics, 405A Atomiştilor Str., 077125 Măgurele-Ilfov, Romania; arpad.rostas@ 123456infim.ro
                [2 ]Department of Inorganic Chemistry, Faculty of Chemistry, University of Bucharest, 90–92 Panduri Str., 050663 Bucharest, Romania; mihaela.badea@ 123456chimie.unibuc.ro (M.B.); catalin.maxim@ 123456chimie.unibuc.ro (C.M.); mirela_luk94@ 123456yahoo.com (M.L.)
                [3 ]Department of Organic Chemistry, Biochemistry and Catalysis, Faculty of Chemistry, University of Bucharest, 90–92 Panduri Str., 050663 Bucharest, Romania; lavinia.ruta@ 123456gmail.com (L.L.R.); ileana.farcasanu@ 123456chimie.unibuc.ro (I.C.F.)
                [4 ]Department of Microbiology, Faculty of Biology, University of Bucharest, 1–3 Aleea Portocalelor Str., 060101 Bucharest, Romania; carmen_balotescu@ 123456yahoo.com (M.C.C.); bmarcelica@ 123456yahoo.com (M.P.)
                [5 ]Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, 1000 Ljubljana, Slovenia; natasa.celan@ 123456fkkt.uni-lj.si (N.C.K.); romana.cerc-korosec@ 123456fkkt.uni-lj.si (R.C.K.)
                [6 ]Department of Life and Environmental Physics, Horia Hulubei National Institute for Physics and Nuclear Engineering, 30 Reactorului Str., 077125 Măgurele-Ilfov, Romania; bmihaela@ 123456nipne.ro (M.B.); mina.raileanu@ 123456nipne.ro (M.R.)
                [7 ]Department of Electricity, Solid State and Biophysics, Faculty of Physics, University of Bucharest, 405A Atomiştilor Str., 077125 Măgurele-Ilfov, Romania
                Author notes
                Author information
                https://orcid.org/0000-0001-8190-9512
                https://orcid.org/0000-0002-4901-7896
                https://orcid.org/0000-0001-6098-1857
                https://orcid.org/0000-0002-3386-6754
                https://orcid.org/0000-0002-1484-5165
                https://orcid.org/0000-0002-7309-6002
                https://orcid.org/0000-0002-4208-0972
                Article
                molecules-25-03777
                10.3390/molecules25173777
                7504215
                32825156
                9d5cc077-fc5a-45a4-83e7-5a825ce6635c
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 13 July 2020
                : 14 August 2020
                Categories
                Article

                copper(ii) complex,1,2,4-triazolo[1,5-a]pyrimidine,cytotoxicity,biofilm,metallonuclease activity,dna intercalation

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