Displacement affinity chromatography of protein phosphatase one (PP1) complexes

Background  

Protein phosphatase one (PP1) is a ubiquitously expressed, highly conserved protein phosphatase that dephosphorylates target protein serine and threonine residues. PP1 is localized to its site of action by interacting with targeting or regulatory proteins, a majority of which contains a primary docking site referred to as the RVXF/W motif.     

Fluorescence and absorption spectroscopic studies on the interaction of porphyrins with snake gourd (Trichosanthes anguina) seed lectin

The interaction of several free-base porphyrins and their corresponding copper(II) and zinc(II) derivatives with the galactose-specific lectin from snake gourd (Trichosanthes anguina) seeds has been investigated by absorption and fluorescence spectroscopic techniques. The lectin dimer contains two apparently equivalent binding sites for the porphyrins. Association constants obtained for the interaction of various porphyrins with the lectin are in the range 1.7 x 10(4)-6.2 x 10(5) M(-1), with the metalloporphyrins being seen to have higher affinity for the lectin compared with the free-base analogues. Both positively charged and negatively charged porphyrins bind to snake gourd seed lectin (SGSL) with comparable affinities, suggesting that binding occurs primarily via hydrophobic interactions. Further, binding of porphyrins is found to be largely unaffected by the presence of the sugar ligand, lactose, indicating that the binding sites for the carbohydrate and porphyrin are different. This study thus suggests that the lectin may serve as a receptor for some endogenous non-carbohydrate, hydrophobic ligand in vivo, in addition to the saccharide ligands. It also opens up the possibility of employing the T. anguina lectin in applications such as photodynamic therapy, which involve the use of porphyrins.     

Schiff bases of nickel(II), copper(II) porphyrins and dibenzo-18-crown-6 interspersed bis -metal porphyrins. Protonation studies

Schiff-base (SB) derivatives of Ni(II) and Cu(II) porphyrins endowed with various amine functions (R−NH₂),n-butylamine,p-anisidine andm-nitroaniline have been prepared from corresponding formyl porphyrins. Protonation studies of these SB derivatives reveal a marked red shift of the optical absorption bands in the visible region relative to the unprotonated imines. The magnitude of the observed red shifts in the protonated derivatives, (SBH⁺) are found to depend on the electron-withdrawing or electron-donating nature of the R group of the amines. The results of the optical absorption,¹H NMR, EPR, and cyclicovoltammetric studies are illustrative of the fact that protonation of the SB derivatives results in a localized positive charge, in the periphery of the porphyrin (p) system. The dibenzo-18-crown-6 interspersedbisporphyrin schiff bases have been prepared fromtrans 4,4′-diamino dibenzo-18-crown-6 and formyl porphyrins. The protonation of these SB derivatives is found to proceed in a concerted fashion. The cation complexation studies by the crown ether entity in thebisporphyrin systems have been investigated using optical absorption, magnetic resonance and electrochemical methods. The redox characteristics of the protonated dimeric SB porphyrins reveal that the first oxidation step involves a two-electron transfer reaction. This is important in view of their possible usage in multielectron transfer reactions of biological and catalytic interest.     

Visual sensing of fluoride ions by dipyrrolyl derivatives bearing electron-withdrawing groups

Two new, easy-to-prepare dipyrrolyl derivatives endowed with electron-withdrawing quinone or dicyano functionalities in their architecture permit the detection of fluoride ions under visual (naked-eye) as well as optical (absorption and fluorescence) and electrochemical conditions in organic solvents     

Chemo- and regioselective cyclohydrocarbonylation of alpha-keto alkynes catalyzed by a zwitterionic rhodium complex and triphenyl phosphite

alpha-Keto alkynes react with CO and H(2) in the presence of catalytic quantities of the zwitterionic rhodium complex (eta(6)-C(6)H(5)BPh(3))(-)Rh(+)(1,5-COD) and triphenyl phosphite affording either the 2-, 2(3H)-, or 2(5H)-furanones in 61-93% yields. The cyclohydrocarbonylation is readily accomplished using substrates containing alkyl, aryl, vinyl, and alkoxy groups at the acetylenic terminal, as well as a variety of primary, secondary, and tertiary alkyl, aryl, and heteroaryl groups connected to the ketone functionality. Structural and electronic properties present in the starting materials mediate the chemo- and regioselectivity of the reaction.     

Ruthenium(II) complexes of 6,7-dicyanodipyridoquinoxaline: synthesis, luminescence studies, and DNA interaction

The hexaflurophosphate and chloride salts of a series of ruthenium(II) complexes incorporating a new dipyridophenazine-based ligand, dicnq (6,7-dicyanodipyrido[2,2-d:2',3'-f]quinoxaline), are synthesized in good-to-moderate yields. These mono ([Ru(phen)2(dicnq)]2+; phen = 1,10-phenanthroline), bis ([Ru(phen)(dicnq)2]2+), and tris ([Ru(dicnq)3]2+) complexes are fully characterized by elemental analysis, infrared, FAB-MS, 1H NMR, and cyclic voltammetric methods. Results of absorption titration and thermal denaturation studies reveal that these complexes are moderately strong binders of calf-thymus (CT) DNA, with their binding constants spanning the range (1-3) x 10(4) M-1. On the other hand, under the identical set of experimental conditions of light and drug dose, the DNA (pBR 322)-photocleavage abilities of these ruthenium(II) complexes follow the order [Ru(phen)2(dicnq)]2+ > [Ru(phen)(dicnq)2]2+ > [Ru(dicnq)3]2+, an order which is the same as that observed for their MLCT emission quantum yields. Steady-state emission studies carried out in nonaqueous solvents and in aqueous media with or without DNA reveal that while [Ru(dicnq)3]2+ is totally nonemissive under these solution conditions, both [Ru(phen)2(dicnq)]2+ and [Ru(phen)(dicnq)2]2+ are luminescent and function as "molecular light switches" for DNA. Successive addition of CT DNA to buffered aqueous solutions containing the latter two complexes results in an enhancement of the emission in each case, with the enhancement factors at saturation being approximately 16 and 8 for [Ru(phen)2(dicnq)]2+ and [Ru(phen)(dicnq)2]2+, respectively. These results are discussed in light of the relationship between the structure-specific deactivations of the MLCT excited states of these metallointercalators and the characteristic features of their DNA interactions, and attempts are made to compare and contrast their properties with those of analogous dipyridophenazine-based complexes, including the ones reported in the preceding paper.     

Ruthenium(II) complexes of redox-related, modified dipyridophenazine ligands: synthesis, characterization, and DNA interaction

The synthesis, spectral characterization, and electrochemical properties of [Ru(phen)2(qdppz)]2+, which incorporates a quinone-fused dipyridophenazine ligand (naphtho[2,3-a]dipyrido[3,2-h:2',3'-f]phenazine-5,18-dione, qdppz), are described in detail. Chemical or electrochemical reduction of [Ru(phen)2(qdppz)]2+ leads to the generation of [Ru(phen)2(hqdppz)](2+)--a complex containing the hydroquinone form (hqdppz = 5,18-dihydroxynaphtho[2,3-a]-dipyrido[3,2-h:2',3'-f]phenazine) of qdppz. Absorption and viscometric titration, thermal denaturation, topoisomerase assay, and differential-pulse voltammetric studies reveal that [Ru(phen)2(qdppz)]2+ is an avid binder of calf-thymus DNA due to a strong intercalation by the ruthenium-bound qdppz, while [Ru(phen)2(hqdppz)]2+ binds to DNA less strongly than the parent "quinone"-containing complex. DNA-photocleavage efficiencies of these complexes also follow a similar trend in that the MLCT-excited state of [Ru(phen)2(qdppz)]2+ is more effective than that of [Ru(phen)2(hqdppz)]2+ in cleaving the supercoiled plasmid pBR 322 DNA (lambda exc = 440 +/- 5 nm), as revealed by the results of agarose gel electrophoresis experiments. The photochemical behaviors of both the quinone- and hydroquinone-appended ruthenium(II) complexes in the presence of DNA not only provide valuable insights into their modes of binding with the duplex but also lead to detailed investigations of their luminescence properties in nonaqueous, aqueous, and aqueous micellar media. On the basis of the results obtained, (i) a photoinduced electron transfer from the MLCT state to the quinone acceptor in Ru(phen)2(qdppz)]2+ and (ii) quenching of the excited states due to proton transfer from water to the dipyridophenazine ligand in both complexes are invoked to rationalize the apparent lack of emission of these redox-related complexes in the DNA medium.     

Redox control of photoinduced electron transfer in axial terpyridoxy porphyrin complexes

The photophysical properties of axial-bonding types (terpyridoxy)aluminum(III) porphyrin (Al(PTP)), bis(terpyridoxy)tin(IV) porphyrin (Sn(PTP) 2), and bis(terpyridoxy)phosphorus(V) porphyrin ([P(PTP) 2] (+)) are reported. Compared with their hydroxy analogues, the fluorescence quantum yields and singlet-state lifetimes were found to be lower for Sn(PTP) 2 and [P(PTP) 2] (+), whereas no difference was observed for Al(PTP). At low temperature, all of the compounds show spin-polarized transient electron paramagnetic resonance (TREPR) spectra that are assigned to the lowest excited triplet state of the porphyrin populated by intersystem crossing. In contrast, at room temperature, a triplet radical-pair spectrum that decays to the porphyrin triplet state with a lifetime of 175 ns is observed for [P(PTP) 2] (+), whereas no spin-polarized TREPR spectrum is found for Sn(PTP) 2 and only the porphyrin triplet populated by intersystem crossing is seen for Al(PTP). These results clarify the role of the internal molecular structure and the reduction potential for electron transfer from the terpyridine ligand to the excited porphyrin. It is argued that the efficiency of this process is dependent on the oxidation state of the metal/metalloid present in the porphyrin and the reorganization energy of the solvent.     

Dose–Response Relationship of Photobiomodulation Therapy on Matrix Metalloproteinase in Healing Dynamics of Diabetic Neuropathic Ulcers—An in vivo Study

Individuals with diabetic foot ulcers have overlapped the inflammatory, proliferative and remodeling phase, making the tissue vulnerable to delayed healing responses. We aimed to establish the dose–response relationship of photobiomodulation therapy of different doses and matrix metalloproteinases in the healing dynamics of diabetic neuropathic ulcers. Diabetes was induced in 126 Albino Wistar rats, and neuropathy was induced to the hind paw by a sciatic nerve injury method. An excisional wound was created on the neuropathy-induced leg. Photobiomodulation therapy of dosages 4, 6, 8, 10, 12 and 15 J cm⁻² and wavelength 655 nm and 808 nm was irradiated. Photobiomodulation therapy of dosages 4, 6 and 8 J cm⁻² showed better wound healing properties with optimized levels of matrix metalloproteinases-1 and 8. We observed a strong dose response in the experimental group treated with 6 and 8 J cm⁻². The findings from the present study conclude that photobiomodulation therapy of dosages 4, 6 and 8 J cm⁻² is suggestive of usefulness in diabetic neuropathic ulcer healing. Markers like matrix metalloproteinases may give a clear direction on response to the therapy. Based on the findings from the present study, we recommend to validate the findings for safety and efficacy in future through human prospective randomized controlled clinical trials.