Biological activity of mixed chelate copper(II) complexes, with substituted diimine and tridentate Schiff bases (NNO) and their hydrogenated derivatives as secondary ligands: Casiopeína's fourth generation.

We synthesize and characterize nine copper(II) compounds. Four with general formula [Cu(NNO)(NO3)] and five mixed chelates [Cu(NNO)(N-N)]+, where NNO corresponds to asymmetric salen ligands (E)-2-((2-(methylamino)ethylimino)methyl)phenolate (L1) and (E)-3-((2-(methylamino)ethylimino)methyl)naphthalenolate (LN1); and their hydrogenated derivatives 2-((2-(methylamino)ethylamino)methyl)phenolate (LH1) and 3-((2-(methylamino)ethylamino)methyl)naphthalenolate (LNH1); and N-N correspond to 4,4'-dimethyl-2,2'-bipiridyne(dmbpy) or 1,10-phenanthroline (phen). Using EPR, the geometries of the compounds in solution in DMSO were assigned, [Cu(LN1)(NO3)] and [Cu(LNH1)(NO3)] a square-planar, [Cu(L1)(NO3)], [Cu(LH1)(NO3)], [Cu(L1)(dmby)]+ and [Cu(LH1)(dmby)]+ a square-based pyramid; and [Cu(LN1)(dmby)]+, [Cu(LNH1)(dmby)]+ and [Cu(L1)(phen)]+ and elongated octahedral. By X-ray it was observed that [Cu(L1)(dmby)]+ and. [Cu(LN1)(dmby)]+ presented a square-based pyramidal, and [Cu(LN1)(NO3)]+ a square-planar geometry. The electrochemical study showed that copper reduction process is a quasi-reversible system, where the complexes with hydrogenated ligands were less oxidizing. The cytotoxicity of the complexes was tested by MTT assay, all the compounds showed biological activity in HeLa cell line, the mixed compounds were the more active ones. Naphthalene moiety, imine hydrogenation and aromatic diimine coordination, increased biological activity. A structure-activity relationships were found: Log(IC50) =  - 1.01(Epc) - 0.35(Conjugated Rings) + 0.87, for Schiff base complexes and Log(IC50) = 0.078(Epc) - 0.32(Conjugated Rings) + 1.94, for hydrogenated complexes; the less oxidizing species with a great number of conjugated rings presented the best biological activity. Complexes-DNA binding constants were obtained by uv-vis studies using CT-DNA, the results suggested that the complexes can interact through the grooves, except the phenanthroline mixed complex that intercalate with DNA. Gel electrophoresis study with pBR 322 showed that compounds can produce changes in the form of DNA and some complexes can cleave DNA in the presence of H2O2.

Structural and functional analysis of a tandem repeat galacturonic acid-binding lectin from the sea hare Aplysia californica.

A d-galacturonic acid-specific lectin, named AcL, was purified from the sea hare Aplysia californica by galactose-agarose affinity chromatography. AcL has a molecular mass of 27.5 kDa determined by MALDI-TOF mass spectrometry. This lectin shows a good affinity for d-galacturonic acid and a lower affinity for galactosides: raffinose, melibiose, α and ß-lactose, and d-galactose. We determined the amino acid sequence of AcL by trypsin digestion and subsequent peptide analysis by mass spectrometry, resulting in a 238 amino acid protein with a theoretical molecular mass of 26.4 kDa. The difference between the theoretical and experimental values can be attributed to post-translational modifications. Thiol-disulfide quantification discerned five disulfide bonds and three free cysteines. The structure of Acl is mainly comprised of beta sheets, determined by circular dichroism, and predicted with AlphaFold. Theoretical models depict three nearly identical tandem domains consisting of two beta sheets each. From docking analysis, we identified AcL glycan-binding sites as multiple conserved motifs in each domain. Furthermore, phylogenetic analysis based on its structure and sequence showed that AcL and its closest homologues (GalULs) form a clear monophyletic group, distinct from other glycan-binding proteins with a jelly-roll fold: lectins of types F and H. GalULs possess four conserved sequence regions that distinguish them and are either ligand-binding motifs or stabilizing network hubs. We suggest that this new family should be referred to as GalUL or D-type, following the traditional naming of lectins; D standing for depilans, the epithet for the species (Aplysia depilans) from which a lectin of this family was first isolated and described.