Investigation of an Allosteric Deoxyhypusine Synthase Inhibitor in P. falciparum

The treatment of a variety of protozoal infections, in particular those causing disabling human diseases, is still hampered by a lack of drugs or increasing resistance to registered drugs. However, in recent years, remarkable progress has been achieved to combat neglected tropical diseases by sequencing the parasites’ genomes or the validation of new targets in the parasites by novel genetic manipulation techniques, leading to loss of function. The novel amino acid hypusine is a posttranslational modification (PTM) that occurs in eukaryotic initiation factor 5A (EIF5A) at a specific lysine residue. This modification occurs by two steps catalyzed by deoxyhypusine synthase (dhs) and deoxyhypusine hydroxylase (DOHH) enzymes. dhs from Plasmodium has been validated as a druggable target by small molecules and reverse genetics. Recently, the synthesis of a series of human dhs inhibitors led to 6-bromo-N-(1H-indol-4yl)-1-benzothiophene-2-carboxamide, a potent allosteric inhibitor with an IC₅₀ value of 0.062 µM. We investigated this allosteric dhs inhibitor in Plasmodium. In vitro P. falciparum growth assays showed weak inhibition activity, with IC₅₀ values of 46.1 µM for the Dd2 strain and 51.5 µM for the 3D7 strain, respectively. The antimalarial activity could not be attributed to the targeting of the Pfdhs gene, as shown by chemogenomic profiling with transgenically modified P. falciparum lines. Moreover, in dose-dependent enzymatic assays with purified recombinant P. falciparum dhs protein, only 45% inhibition was observed at an inhibitor dose of 0.4 µM. These data are in agreement with a homology-modeled Pfdhs, suggesting significant structural differences in the allosteric site between the human and parasite enzymes. Virtual screening of the allosteric database identified candidate ligand binding to novel binding pockets identified in P. falciparum dhs, which might foster the development of parasite-specific inhibitors.     

基于变构学习模型的高中化学“导致悖论”教学设计*

“导致悖论”教学是一种智在创境、妙在引领的教学,其核心理念源自于变构学习模型的悖论思想,给学习者创造一个“肯定→否定→肯定”的反思性学习过程。高中化学教学设计可以通过4种途径分别创设“概念与概念”相悖、“理论与实验”相悖、“理论与理论”相悖和“实验与实验”相悖的悖论情境,最终实现学生新旧概念的更替和高阶思维能力的发展。“导致悖论”教学在实施中要注意精选合适的知识内容,加强学生心理的调控,提供丰富的支撑材料。     

Allosteric Modulation of Human Serum Albumin Induced by Peptide Ligand

Human serum albumin (HSA ) is an abundant protein in plasma that can bind and transport many small molecules, and the corresponding affinity‐controlled drug delivery shows great advantage in the biological system. Peptide SA06 is a reported ligand comprising 20 amino acids, and is known to non‐covalently bind with HSA to extend the lifetime and improve the pharmacokinetic performance. The structural information of the HSA ‐peptide complex is keen for obtaining molecular insight of the binding mechanism. We studied the secondary structural change and structure‐affinity relations of Peptide SA06 with HSA by using circular dichroism (CD ) spectroscopy in solution. Noticeable allosteric effect can be identified by compositional increase of α ‐helix structures when the peptide was co‐incubated with HSA . Furthermore, the equilibrium dissociation constant of Peptide SA06 with HSA can be determined by CD ‐based method. This work provides structural evidence on the allosteric interaction between peptide ligand and HSA , and sheds light on optimization of therapeutic properties in the affinity‐controlled delivery systems.     

Current Knowledge and Perspectives of Pyrrolizidine Alkaloids in Pharmacological Applications: A Mini-Review

Pyrrolizidine alkaloids (PAs) are a widespread group of secondary metabolites in plants. PAs are notorious for their acute hepatotoxicity, genotoxicity and neurological damage to humans and animals. In recent decades, the application of PAs for beneficial biological activities to cure disease has drawn greater attention. Here, we review the current knowledge regarding the pharmacological properties of PAs and discuss PAs as promising prototypes for the development of new drugs.     

Nanosecond Dynamics Regulate the MIF‐Induced Activity of CD74

Macrophage migration inhibitory factor (MIF) activates CD74, which leads to severe disorders including inflammation, autoimmune diseases and cancer under pathological conditions. Molecular dynamics (MD) simulations up to one microsecond revealed dynamical correlation between a residue located at the opening of one end of the MIF solvent channel, previously thought to be a consequence of homotrimerization, and residues in a distal region responsible for CD74 activation. Experiments verified the allosteric regulatory site and identified a pathway to this site via the MIF β‐strands. The reported findings provide fundamental insights on a dynamic mechanism that controls the MIF‐induced activation of CD74.     

A Metalloregulated Four‐State Nanoswitch Controls Two‐Step Sequential Catalysis in an Eleven‐Component System

The nanomechanical switch 1 with its three orthogonal binding motifs—the zinc(II) porphyrin, azaterpyridine, and shielded phenanthroline binding station—is quantitatively and reversibly toggled back and forth between four different switching states by means of addition and removal of appropriate metal‐ion inputs. Two of the four switching stages are able to initiate catalytic transformations (ON1, ON2), while the two others shut down any reaction (OFF1, OFF2). Thus, in a cyclic four‐state switching process the sequential transformation A + B + C → AB + C → ABC can be controlled, which proceeds stepwise along the switching states OFF1→ON1 (click reaction: A + B → AB )→OFF2→ON2 (Michael addition: AB + C → ABC )→OFF1. Two consecutive cycles of the sequential catalysis were realized without loss in activity in a reaction system with eleven different components.     

Functional supramolecular systems with highly cooperative and responding properties

The dynamic processes of host–guest interactions contribute to the multistep regulation of various molecular functions such as the catalysis of chemical reactions, transport of materials, control of reaction pathways, and cooperative and responsive phenomena particularly in biological systems. In this review, we describe artificial metallo‐supramolecular systems, which exhibit highly cooperative and responsive functions to external stimulus, by utilizing formation of the metal complexes and their characteristic properties. Pseudomacrocycles such as pseudocrown ethers and pseudocryptands have been synthesized to control macrocyclic and macrobicyclic effects on guest recognition by using a metal ion as an effector, and remarkably positive and negative allosteric effects have been achieved. Highly cooperative stepwise regulation of the affinity to anions has also been achieved by a pseudocryptand and a salt‐binding host. The electrostatic interactions between the anions and cations are important for the combination specificity of the salts. We also introduce a linear bis‐salamo ligand as a precursor for a novel multimetal cooperative host. A trinuclear zinc complex was formed cooperatively and only the central zinc ion was replaced by lanthanide and calcium ions in a transmetalation way. © 2008 The Japan Chemical Journal Forum and Wiley Periodicals, Inc. Chem Rec 8: 240–251; 2008: Published online in Wiley InterScience ( www.interscience.wiley.com ) DOI 10.1002/tcr.20153