Inhibition of brain enolases by acrylamide and its related compounds in vitro,and the structure-activity relationship

Summary Acrylamide and its related compounds inhibited brain enolases in vitro independently of their neurotoxicity. The inhibitory potency was a function of the binding constants of the compounds for phenylalanine. The binding constant for tryptophan was higher in neurotoxic compounds than in non-neurotoxic ones.     

Stereoselectivity of a radioimmunoassay for the insecticide S-bioallethrin

The stereoselectivity of a radioimmunoassay (RIA) system using an S-bioallethrin specific antiserum was studied by observing the abilities of the 8 allethrin isomers and other selected compounds to compete with a radiolabelled S-bioallethrin tyramine derivative for antibody binding sites. The results demonstrate the feasibility of RIA as a rapid, sensitive and steroselective residue technique for compounds difficult to analyze by classical methods.     

Stereoselectivity of a radioimmunoassay for the insecticide S-bioallethrin

Summary The stereoselectivity of a radioimmunoassay (RIA) system using an S-bioallethrin specific antiserum was studied by observing the abilities of the 8 allethrin isomers and other selected compounds to compete with a radiolabelled S-bioallethrin tyramine derivative for antibody binding sites. The results demonstrate the feasibility of RIA as a rapid, sensitive and stereoselective residue technique for compounds difficult to analyze by classical methods.This work was supported in part by grant No. 5-R01-ES01260-02 from the National Institutes of Health.     

Anti-muscarinic activity of a family of C11N5 compounds isolated from Agelas sponges.

In a search for potential target sites for C11N5 compounds obtained from marine sponges of the genus Agelas we evaluated their interaction with muscarinic acetylcholine receptors from rat brain membranes. In competition experiments with 3H-QNB these compounds displayed the following rank order of potency: sceptrin greater than oroidin greater than or equal to dibromosceptrin greater than or equal to clathrodin. Sceptrin (50 microM) was shown to be a competitive inhibitor of 3H-QNB binding as revealed by Scatchard analysis. The results demonstrate the ability of these compounds to interact with multiple target molecules in the micromolar range.     

Effect of lipophilic cations on thiamine transport system in isolated rat hepatocytes

The effect of lipophilic cations such as triphenylmethylphosphonium, tetraphenylphosphonium and tetraphenylarsonium in addition to dibenzyldimethylammonium on thiamine transport in isolated rat hepatocytes was studied. Lipophilic cations at the concentration 10 microM almost completely inhibited thiamine uptake. Kinetic studies showed that these compounds were competitive inhibitors with a very high affinity. These results suggest that lipophilic cations in addition to quaternary ammonium compounds also share a common binding site for thiamine in isolated rat hepatocytes.     

交叉学科研究推动了生物无机化学学科的发展

本文分为七部分，第一部分对生物无机化学学科发展进行历史回顾；第二部分从回顾史实中用实例表明生物无机化学的研究始终瞄准金属离子和生物配体控制生命过程中的化学问题；第三部分是介绍当今国际上研究核酸领域的争论焦点；第四部分是报导我们设计和合成了作为人工核酸酶的一系列新的配体以及许多不同金属的功能配合物；第五部分是讨论用动力学、热力学和DFT计算方法研究配合物和DNA相互作用的键合机制以及十多种影响因素；第六部分进一步报导配合物的生物功能和它们的应用探索，最后一部分提出在核酸领域的某些新的发展方向和途径。我们进一步提出了配合物的结构与DNA的作用机制以及它们的生物功能之间的规律性。通过配合物的结构改变去调控和改变配合物对DNA键合性质和生物功能。     

Enrichment of ligands with molecular dockings and subsequent characterization for human alcohol dehydrogenase 3

Alcohol dehydrogenase 3 (ADH3) has been assigned a role in nitric oxide homeostasis due to its function as an S-nitrosoglutathione reductase. As altered S-nitrosoglutathione levels are often associated with disease, compounds that modulate ADH3 activity might be of therapeutic interest. We performed a virtual screening with molecular dockings of more than 40,000 compounds into the active site of human ADH3. A novel knowledge-based scoring method was used to rank compounds, and several compounds that were not known to interact with ADH3 were tested in vitro. Two of these showed substrate activity (9-decen-1-ol and dodecyltetraglycol), where calculated binding scoring energies correlated well with the logarithm of the k _cat/K _m values for the substrates. Two compounds showed inhibition capacity (deoxycholic acid and doxorubicin), and with these data three different lines for specific inhibitors for ADH3 are suggested: fatty acids, glutathione analogs, and cholic acids.     

Effects of dosage and cadmium pretreatment on the binding of cadmium in rat bile.

The effects of dosage and of cadmium pretreatment on the binding of cadmium in rat bile were studied. With increasing dose a higher cumulative biliary excretion of Cd was observed and a higher percentage of the Cd was excreted in a low-molecular-weight form. On the other hand, after cadmium pretreatment, a decrease in the cumulative biliary excretion of cadmium was observed but a greater percentage of that excreted into the bile was bound to high molecular weight compounds.     

Effects of dosage and cadmium pretreatment on the binding of cadmium in rat bile

Summary The effects of dosage and of cadmium pretreatment on the binding of cadmium in rat bile were studied. With increasing dose a higher cumulative biliary excretion of Cd was observed and a higher percentage of the Cd was excreted in a low-molecular-weight form. On the other hand, after cadmium pretreatment, a decrease in the cumulative biliary excretion of cadmium was observed but a greater percentage of that excreted into the bile was bound to high molecular weight compounds.     

Über die Beeinträchtigung der Wirkung von Spindelgiften in der Zellkultur durch menschliches Serum sowie durch Seren verschiedener Tierspezies

Summary Chick embryo fibroblasts, cultivatedin vitro, require higher concentrations of podophyllotoxin derivatives for complete mitotic arrest when the medium contains human adult or cord serum than when it contains horse, bovine, guinea pig, rat, mouse, or cock serum. This difference is due to a higher binding capacity of human serum for these compounds. No such difference was found with a colchicine and a peltatin derivative.     

Displacement activity of some natural cularine alkaloids at striatal3H-SCH 23 390 and3H-raclopride binding sites

Five natural cularines isolated from the aerial parts ofSarcocapnos crassifolia (Fumariaceae) and a cularioid isolated from the bark ofGuatteria ouregou (Annonaceae) were tested for their ability to displace³H-SCH 23 390 and³H-raclopride from their striatal binding sites. Celtisine, breoganine and cularidine were able to inhibit the binding at D-1 and D-2 dopaminergic sites at nanomolar concentrations. Other alkaloids were active at micromolar concentrations. These data suggest that the presence of an oxepine system in the isoquinoline skeleton could lead to compounds which would be very active and possibly selective at dopaminergic receptor sites.     

Methods of probing the interactions between small molecules and disordered proteins

It is generally recognized that a large fraction of the human proteome is made up of proteins that remain disordered in their native states. Despite the fact that such proteins play key biological roles and are involved in many major human diseases, they still represent challenging targets for drug discovery. A major bottleneck for the identification of compounds capable of interacting with these proteins and modulating their disease-promoting behaviour is the development of effective techniques to probe such interactions. The difficulties in carrying out binding measurements have resulted in a poor understanding of the mechanisms underlying these interactions. In order to facilitate further methodological advances, here we review the most commonly used techniques to probe three types of interactions involving small molecules: (1) those that disrupt functional interactions between disordered proteins; (2) those that inhibit the aberrant aggregation of disordered proteins, and (3) those that lead to binding disordered proteins in their monomeric states. In discussing these techniques, we also point out directions for future developments.     

Proton motive force mediates a reorientation of the cytosolic domains of the multidrug transporter LmrP

LmrP from Lactococcus lactis is a 45-kDa membrane protein that confers resistance to a wide variety of lipophilic compounds by acting as a proton motive force-driven efflux pump. This study shows that both the proton motive force and ligand interaction alter the accessibility of cytosolic tryptophan residues to a hydrophilic quencher. The proton motive force mediates an increase of LmrP accessibility toward the external medium and results in higher drug binding. Residues Asp128 and Asp68, from cytosolic loops, are involved in the proton motive force-mediated accessibility change. Ligand binding does not modify the protein accessibility, but the proton motive force-mediated restructuring is prerequisite for a subsequent accessibility change mediated by ligand binding. Asp142 cooperates with other membrane-embedded carboxylic residues to promote a conformational change that increases LmrP accessibility toward the hydrophilic quencher. This drug binding-mediated reorganization may be related to the transition between the high- and low-affinity drug-binding sites and is crucial for drug release in the extracellular medium.     

Anti-muscarinic activity of a family of C11N5 compounds isolated fromAgelas sponges

In a search for potential target sites for C₁₁N₅ compounds obtained from marine sponges of the genusAgelas we evaluated their interaction with muscarinic acetylcholine receptors from rat brain membranes. In competition experiments with³H-QNB these compounds displayed the following rank order of potency: sceptrin>oroidindibromosceptrinclathrodin. Sceptrin (50 M) was shown to be a competitive inhibitor of³H-QNB binding as revealed by Scatchard analysis. The results demonstrate the ability of these compounds to interact with multiple target molecules in the micromolar range.     

Oxazolidinones: a novel class of antibiotics

Oxazolidinones are a novel class of synthetic antimicrobial agents which have now entered phase III clinical trials. The most promising feature of these compounds is their oral activity against multidrug-resistant Gram-positive bacteria which have created tremendous therapeutic problems in recent years. In addition, development of resistance in vitro has so far remained below detectable levels. Different from many antibacterial agents used in the treatment of human infections, oxazolidinones do not block bacterial protein synthesis at the level of polypeptide chain elongation but rather seem to interfere with initiation of translation. Both binding of formylmethionine-transfer RNA to initiation complexes as well as release of formylmethioninepuromycin from initiation complexes have been reported to be targets for oxazolidinones. The major binding sites of oxazolidinones are the large (50S) ribosomal subunits.     

Modulation of dopaminergic transmission by alpha-noradrenergic agonists and antagonists: Evidence for antidopaminergic properties of some alpha antagonists

Summary The effects on dopamine (DA) metabolism, on³H-spiperone binding and on amphetamine-induced stereotypies of a variety of drugs with different actions on alpha₁-and alpha₂-noradrenergic (NA) receptors have been investigated.The preferential alpha₂-antagonists yohimbine, rauwolscine, piperoxane and esproquin as well as the preferential alpha₁-antagonists corynanthine and WB4101 increased homovanillic acid (HVA) and 3,4-dihydroxyphenylacetic acid (DOPAC) in the rat striatum, mesolimbic area, and cortex. Prazosine and clonidine tended to reduce HVA and DOPAC. The preferential alpha₂-antagonists, tolazoline and RX-781094A, had no measurable effects on DA metabolism even at high doses.Those compounds which in comparable doses increased DA metabolism inhibited³H-spiperone binding in the hippocampus. The effects in the striatum and cortex were smaller and did not show a relation to those in hippocampus or on DA metabolism. Only the yohimbine alkaloids antagonized amphetamine-induced stereotypies.The results suggest that the effects on DA metabolism at least of yohimbine, rauwolscine, and corynanthine are related to their intrinsic antidopaminergic properties. The same might be true, although with a lesser degree of certainty, for piperoxane, esproquin, and WB4101.Since many of the tested compounds possessing alpha-antagonistic properties interacted with the DA system, a close molecular relationship between alpha-noradrenergic and DA receptors might be anticipated. The preference of these compounds for the hippocampal subtype of DA receptors might indicate a particular role of the latter in the regulation of DA metabolism. On the other hand, the antagonism against haloperidol's enhancing effect on DA metabolism by clonidine suggests a modulatory NA influence on DA transmission. The observation that clonidine reduced the effects of yohimbine and piperoxane to a lesser degree than that of haloperidol, is in agreement with this notion.Part of this work has been presented at the 13th Meeting of the Union of Swiss Societies of Experimental Biology, Lausanne, March 26/27. 1981 (for abstract see Waldmeier and Bischoff, 1981).     

Cooperative interactions between odorant-binding proteins of Anopheles gambiae

To understand olfactory discrimination in Anopheles gambiae, we made six purified recombinant OBPs and investigated their ligand-binding properties. All OBPs were expressed in bacteria with additional production of OBP47 in the yeast Kluveromyces lactis. Ligand-binding experiments, performed with a diverse set of organic compounds, revealed marked differences between the OBPs. Using the fluorescent probe N-phenyl-1-naphthylamine, we also measured the binding curves for binary mixtures of OBPs and obtained, in some cases, unexpected behaviour, which could only be explained by the OBPs forming heterodimers with binding characteristics different from those of the component proteins. This shows that OBPs in mosquitoes can form complexes with novel ligand specificities, thus amplifying the repertoire of OBPs and the number of semiochemicals that can be discriminated. Confirmation of the likely role of heterodimers was demonstrated by in situ hybridisation, suggesting that OBP1 and OBP4 are co-expressed in some antennal sensilla of A. gambiae.     

The pleiotropic functions of aspirin: mechanisms of action

Recent studies have suggested that aspirin and aspirin-like compounds have a variety of actions in addition to their well-studied ability to inhibit cyclooxygenases. These actions include inhibition of the uncoupling of oxidative phosphorylation, decreases in adenosine triphosphate stores, increases in extracellular adenosine, downregulation of the expression and activity of inducible nitric oxide synthetase, inhibition and/or stimulation of various mitogen-activated protein kinase activities and inhibition of nuclear factor binding κB site (NF-κB) activation. Moreover, aspirin-like compounds have recently been shown to have previously unappreciated clinical and biological effects, some apparently independent of cyclooxygenase. In this review we discuss the various mechanisms of action of aspirin-like compounds and their relevance to clinical disease and therapy. Received 1 February 1999; received after revision 1 April 1999; accepted 7 May 1999     

Complexing agents from microorganisms

Summary The majority of extracellular complexing ligands produced by microorganisms are summarized as being of low molecular mass (<10,000 daltons) and are usually released as part of metal detoxification processes. These exudates appear to exhibit strong metal-binding characteristics, often reducing metal toxicity. Under certain conditions microbes produce metal-specific compounds of low molecular mass called siderophores; although these are normally specific for iron they also have relatively high affinities for radionuclides such as Pu and facilitate their uptake into cells. The occurrence of specific actinide complexing agents has been recorded.The breakdown of lignins and cellulosic material produces large macromolecular compounds called humates. These contain multiligand sites and display a wide range of complexing abilities. They form both soluble and insoluble complexes with toxic elements with various results. Humates also considerably influence adsorption of metals to substrate surfaces and at high pH may compete with OH-ions for metal binding.As well as with extracellular ligands, metals can interact directly with microorganisms by accumulation in subcellular compartments or by adsorption on bacterial surfaces.     

Extraction and purification of brain glycoprotein NSA3 by binding with hyaluronic acid

Brain glycoprotein NSA3 was found to bind to immobilised hyaluronic acid. The binding was reversible and gave pure antigen (99%) in high yields (80%). The binding was suppressed by incubating the affinosorbent with hyaluronidase. It was not suppressed by trypsin. The presence of glycosaminoglycans other than hyaluronic acid in the sample did not inhibit the binding. This property is relevant to those already known for the brain glycoprotein NSA3, and to its localisation at the nodes of Ranvier. We propose to coin the name hyaluronectin for the brain glycoprotein NSA3.