Insight into the interaction of benzothiazole tethered triazole analogues with human serum albumin: Spectroscopy and molecular docking approaches

The interaction of four benzothiazole tethered triazole analogues (MS43, MS70, MS71, and MS78) with human serum albumin (HSA) was investigated using various spectroscopic techniques (ultraviolet–visible (UV–vis) light absorption, fluorescence, circular dichroism (CD), molecular docking and density functional theory (DFT) studies). Fluorescence quenching constants (~10¹²) revealed a static mode of quenching and binding constants (K_b ~10⁴) indicating the strong affinity of these analogues for HSA. Further alteration in the secondary structure of HSA in the presence of these analogues was also confirmed by far UV–CD spectroscopy. The intensity loss in CD studied at 222 nm indicated an increase in random coil/β‐sheet conformations in the protein. Binding energy values (MS71 (?9.3 kcal mol^?1), MS78 (?8.02 kcal mol^?1), MS70 (?7.16 kcal mol^?1) and MS43 (?6.81 kcal mol^?1)) obtained from molecular docking revealed binding of these analogues with HSA. Molecular docking and DFT studies validated the experimental results, as these four analogues bind with HSA at site II through hydrogen bonding and hydrophobic interactions.     

Synthesis of spiro[chroman-2,4′-piperidin]-4-one derivatives as acetyl-CoA carboxylase inhibitors

Various spiro[chroman-2,4′-piperidin]-4-one derivatives (38a–m and 43a–j) have been designed, synthesized and evaluated for in vitro acetyl-CoA carboxylase (ACC) inhibitory activity. Several compounds have shown ACC inhibitory activity in low nanomolar range. Compound 38j reduced the respiratory quotient (RQ) in C57BL/6J mice indicating increase in whole body fat oxidation even in the presence of high carbohydrate diet. Structure–activity relationship (SAR) has been discussed.     

Assessment of membrane damage in continuous cultures of mammalian cells

The present studies were aimed at evaluating procedures for assessing the effect of chemicals on the integrity of the plasma membrane in continuous cell cultures. The degree of membrane damage was monitored by determining the ‘leakage’ of α-[³H]aminoisobutyric acid ([³H]AIB) and [¹⁴C]deoxy-2-fluoro-D-glucose ([¹⁴C]FdG) from the prelabelled cells. These parameters were compared to the loss of lactate dehydrogenase (LDH) from the cells and the decrease in the intracellular level of K⁺. Triton X-100, sodium dodecylsulfate (SDS), phospholipase C and nystatin which are known to affect membranes by different mechanisms served as test agents. In parallel, we monitored the effects of the chemicals on the viability of the cells. The following results were obtained:(1) The two radioactive markers [³H]AIB and [¹⁴C]FdG were found to be suitable to probe for damages of the plasma membrane in a variety of continuous cell lines which differ widely in their phenotype, rate of growth and degree of differentiation. (2) The leakage of the two markers could conveniently be monitored by double labelling techniques. (3) The loss from the cells of the 3 markers of smaller molecular size, K⁺, [³H]AIB, [¹⁴C] FdG, differed considerably depending on the test agent used. (4) Intracellular K⁺ level and [³H]AIB leakage generally appeared to follow a similar pattern, whereas [¹⁴C]FdG leakage may have shown a distinctly different response. (5) The leakage of LDH was an insensitive indicator for membrane damage. (6) No clear relationship was detectable between a particular leakage pattern of the markers and the loss of cellular viability.     

S-100-Mediated Inhibition of Brain Protein Phosphorylation

The effects of the glial-specific, calcium-binding, S-100 protein on brain membrane and supernatant protein phosphorylation were assessed. S-100 concentrations as low as 5 micrograms/ml caused a marked inhibition of the phosphorylation of a soluble brain protein having a molecular weight of 73,000 daltons (73K). This protein was designated the S-100 protein-modulated phosphoprotein (SMP). Half-maximal inhibition of the phosphorylation of SMP by S-100 was obtained at concentrations of 12 micrograms/ml (0.57 microM). The inhibition of SMP phosphorylation by S-100 was calcium-dependent, with a calculated calcium Ka of 2.0 +/- 0.3 microM. SMP phosphorylation was also inhibited by calmodulin, but only partially and with a much lower potency. The inhibition of SMP phosphorylation by S-100 was not inhibited by fluphenazine, whereas the effect of calmodulin was. SMP was found in many brain areas, with the highest levels seen in the corpus callosum. Various peripheral tissues, such as kidney; liver; and pineal, pituitary, and adrenal glands, did not contain detectable SMP levels. At higher S-100 concentrations, greater than 10 micrograms/ml, the phosphorylation of several other soluble proteins was markedly inhibited. These proteins have molecular weights of 56K, 50K, and 47K. The phosphorylation of these proteins was enhanced by calmodulin. These data suggest that the S-100 protein may function to modulate the phosphorylation of brain proteins in a manner analogous to (although in a reciprocal fashion) that of calmodulin.     

Differential Binding Properties of Adenosine Receptor Agonists and Antagonists in Brain

The binding properties of N6-cyclohexyl [3H]adenosine ( [3H]CHA) and 1,3-diethyl-8-[3H]phenylxanthine ( [3H]DPX) in rat forebrain membrane are compared. The kinetic parameters of binding for each ligand are quite distinct, with [3H]CHA displaying two populations of binding sites (KD = 0.4 +/- 0.05 nM and 4.2 +/- 0.3 nM; Bmax = 159 +/- 17 and 326 +/- 21 fmol/mg protein), whereas [3H]DPX yielded monophasic Scatchard plots (KD = 13.9 +/- 1.1 nM; Bmax = 634 +/- 27 fmol/mg protein). The metals copper, zinc, and cadmium are potent inhibitors of [3H]CHA binding, with respective IC50 concentrations of 36 microM, 250 microM, and 70 microM. Copper is a much less potent inhibitor of [3H]DPX binding (IC50 = 350 microM). The inhibitory effect of copper on both [3H]CHA and [3H]DPX binding is apparently irreversible, as membranes pretreated with copper cannot be washed free of its inhibitory effect. The inhibitory effect of both copper and zinc on [3H]CHA binding was reversed by the guanine nucleotide Gpp(NH)p. [3H]DPX binding is only partially inhibited by zinc and cadmium (60% of specific binding remains unaffected), suggesting that this adenosine receptor ligand binds to two separate sites. Guanine nucleotides had no effect on the inhibition of [3H]DPX binding by either copper or zinc. Differential thermal and proteolytic denaturation profiles are also observed for [3H]CHA and [3H]DPX binding, with the former ligand binding site being more labile in both cases. Stereospecificity is observed in the inhibition of both [3H]CHA and [3H]DPX binding, with L-N-phenylisopropyladenosine (PIA) being 50-fold more potent than D-PIA in both cases. Evidence is therefore provided that adenosine receptor agonists and antagonists have markedly different binding properties to brain adenosine receptors.     

Cyclic AMP-binding capacities and histone kinase activation in subcellular components of neocortical tissue. Differential responses to three neurohumoural agents

1. Noradrenaline and histamine, when added to superfused guinea-pig cerebral-cortical tissues, increased both cyclic AMP-dependent and -independent histone kinase activities of some, but not of all, subsequently isolated subcellular fractions, and decreased their cyclic [(3)H]AMP-binding capacity, which was concluded to be due to an increase in endogenously bound cyclic AMP. 2. Adenosine and 2-chloroadenosine also diminished the cyclic [(3)H]AMP-binding capacities, but did not affect the histone kinase activities. 3. DEAE-cellulose chromatography and stability to KCl additions showed that the greater part of the histone kinase of the present preparations corresponded to the type II enzyme [of Corbin, Keely & Park (1975) J. Biol. Chem.250, 218-225], with a lesser amount of type I activity. Different sites of cyclic AMP accumulation in relation to these or other kinases are considered in interpreting the differential tissue responses to the neurohumoural agents examined.     

Some effects of salicylic acid on growth and flowering in Spirodela polyrrhiza SP20

More than 70% flowering was observed in Spirodela polyrrhizaSP₂₀ under long and short days, when fronds were grown in amedium supplemented with 5x10^–5 M salicylate (SA). Toour knowledge this is the first report of such profuse floweringof this duckweed in vitro. Besides flowering, SA also affectedthe multiplication rate, anthocyanin and chlorophyll contents,and gibbosity. Aspirin had an effect similar to that of SA. (Received April 1, 1980; )