Nmr and Mass Spectroscopic Studies of the Competitive-Angiotensin II Antagonist “Sarmesin”

Fast atom bombardment mass spectrometry (FAB-MS) and high resolution (400 Mz) proton nuclear magnetic resonance (NMR spectroscopy) on the competitive angiotensin II antagonist, |Sar¹, Tyr(Me)⁴ (ANGII (Sarmesin) and its he-ptapeptide homolog, [Tyr(Me)³ |ANGIII, yield spectra which provide confirmation of structure and molecular weight. The characteristics of the spectra are discussed and compared with the spectra of natural ANG II, ANG III and |Sar¹|ANG II. The NMR data are suggestive of interactions in angiotensin between: 1) the phenolic hydroxyl group and the imidazole ring, and 2) the N-terminal amino group and the Tyrring. These interactions may be important for the formation of the proposed charge transfer system in angiotensin II involving the phenoxyl and α-carboxylate groups.     

Synthesis,bioactivity and functional evaluation of linker-modified allatostatin analogs as potential insect growth regulators

Insect growth regulators play an important role in integrated pest management strategies.The FGLa–allatostatins(ASTs)are a family of neuropeptides that can inhibit juvenile hormone(JH)biosynthesis by the corpora allata(CA)of Diploptera punctata in vitro,are regarded as insect growth regulator candidates.In the search for new potential mimics and to explore the effect of linker length on inhibiting JH biosynthesis,a series of AST analogs were synthesized by modifying the linker of K24,which was found to have a significant effect on JH biosynthesis in vitro in our previous study.Functional evaluation demonstrated that all the target compounds can activate the Dippu-Ast R,albeit with different potencies.Analog L6 with the longest linker(n=5),exhibited not only a promising effect on inhibition of JH biosynthesis both in vitro and in vivo,but also good activity in inhibiting basal oocyte growth.Structure–activity relationships(SAR)studies showed that longer linkers provided greater contribution to activity.     

Characterization of Compounds Acting on the α1A Adrenergic Receptor from Caulis spatholobi by Cell Membrane Chromatography with Possible Application for Treatment of Benign Prostatic Hyperplasia

Tamsulosin hydrochloride is a commonly used drug for treatment of benign prostatic hyperplasia which is a disease affecting elderly men. However, adverse reactions may occur when tamsulosin hydrochloride is used for long-term treatment. Caulis spatholobi is a traditional Chinese medicine with many pharmacological effects. In this study, a cell membrane chromatography column was combined with high-performance liquid chromatography–mass spectrometry to determine the active compounds acting on α1A adrenergic receptor from Caulis spatholobi. Formononetin was shown to be active on the α1A adrenergic receptor. This was investigated by a competitive binding assay that demonstrated that tamsulosin hydrochloride and formononetin employ the same binding sites. Thus, formononetin is a potential α1A adrenergic receptor antagonist for treating benign prostatic hyperplasia.     

In vitro bioassays for the study of endocrine-disrupting food additives and contaminants

Endocrine-disrupting chemicals (EDCs) are capable of interfering with normal hormone homeostasis by acting on several targets and through a wide variety of mechanisms. Unwanted exposure to EDCs can lead to a wide spectrum of adverse health effects, especially when exposure is during critical windows of development. Feed and food are considered to be among the main routes of inadvertent exposure to EDCs, so there is an important need for efficient detection of EDCs in these matrices.We describe in vitro bioassays that can complement current analytical chemistry in order to detect unwanted EDCs and describe their action, emphasizing assays that can measure effects on nuclear receptor signaling or hormone production. We outline both validated and unvalidated in vitro assays currently available in the scientific community for detecting and studying the effects of EDCs, and discuss their possible role in the food-safety context. We conclude by identifying gaps in the current battery of in vitro assays available for EDCs and suggest future possibilities for development and validation.     

Progress in DNA Aptamers as Recognition Components for Protein Functional Regulation

Proteins play a central role in all domains of life, and precise regulation of their activity is essential for understanding the related biological processes and therapeutic functions. Nucleic acid aptamers, the molecular recognition components derived from systematic evolution of ligands by exponential enrichment(SELEX), can specifically identify proteins with antibody-like recognition characteristics and help to regulate their activity. This minireview covers the SELEX-based selection of protein-binding aptamers, membrane protein analytical techniques based on aptamer-mediated target recognition, aptamer-mediated functional regulation of proteins, including membrane receptors and non-membrane proteins(thrombin as a model), as well as the potential challenges and prospects regarding aptamer-mediated protein manipulation, aiming to supply some useful information for researchers in this field.     

Photoaffinity Probe Candidates for Gamma-aminobutyric Acid （GABAA）-Gated Chloride Channel

New photoaffinity ligand candidates were synthesized based on 5-t-buty1-2-(4-(substituted-ethyny1)phenyl)-1, 3-dithiane for the noncompetitive blocker site on the gammaaminobutyric acid-gated chloride channel. Their half-maximal inhibition concentrations ranged from 4 to 32 nmol/L as measured by 4‘-ethyny1-4-n-[2,3-3^H2]-propylbicycloorthobenzoate (3^H EBOB) assay.     

Membrane Organization and Dynamics of the G-Protein-Coupled Serotonin<Subscript>1A</Subscript> Receptor Monitored Using Fluorescence-Based Approaches

The G-protein-coupled receptor (GPCR) superfamily represents one of the largest classes of molecules involved in signal transduction across the plasma membrane. Fluorescence-based approaches have provided valuable insights into GPCR functions such as receptor–receptor and receptor–ligand interactions, real-time assessment of signal transduction, receptor dynamics on the plasma membrane, and intracellular trafficking of receptors. This has largely been possible with the use of fluorescent probes such as the green fluorescent protein (GFP) from the jellyfish Aequoria victoria and its variants. We discuss the potential of fluorescence-based approaches in providing novel information on the membrane organization and dynamics of the G-protein-coupled serotonin_1A receptor tagged to the enhanced yellow fluorescent protein (EYFP). These authors contributed equally to the work.     

Discovery of structurally diverse natural product antagonists of chemokine receptor CXCR3

The chemokines (CXCL9, CXCL10 and CXCL11) and associated CXCR3 receptor are expressed during the inflammatory process from multiple sclerosis, atherosclerosis or organ transplantation resulting in the recruitment of lymphocytes leading to tissue damage. It is hypothesized that blocking of the ligand/CXCR3 receptor interaction has potential to provide opportunity for development of agents that would block tissue rejection. In this paper, four classes of natural product inhibitors (IC₅₀ ranging 0.1–41 M) have been described that block the CXCR3 receptor interaction of IP-10 ligand. These include a cyclic thiopeptide (duramycin), polyketide glycosides (roselipins), steroidal glycosides (hypoglausin A and dioscin) and a novel alkyl pyridinium alkaloid that were isolated by bioassay-guided fractionation of the organic extracts derived from actinomycete, fungal, plant and marine sources and discovered using ¹²⁵ I IP-10/CXCR3 binding assay. Duramycin was the most potent with an IC₅₀ of 0.1 M. Roselipins 2A, 2B and 1A showed IC₅₀ values of 14.6, 23.5, and 41 M, respectively. Diosgenin glycosides dioscin, hypoglaucin A and kallstroemin D exhibited IC₅₀ values of 2.1, 0.47 and 3 M, respectively. A novel cyclic 3-alkyl pyridinium salt isolated from a sponge displayed a binding IC₅₀ of 0.67 M.     

A functionalized dianthryl tetraaza macrocycle having the recognizing and switching ability

The synthesized macrocycle L is the novel fluorescent receptor having the switching ability by the external stimuli as well as having the recognizing ability of various metal ions. In particular, this macrocycle L shows the possibility of the selectivity of metal ions even in the same charge ions of a different metal, and the values of association constant (M⁻¹) of that for metal ions are consistent with the tendency of increasing charge number of metal ion. In addition, the values of quantum yield (Φ_F) of metal complexes of macrocycle L were ranged from 0.021 to 0.111 enough to recognize the metal ions in macrocycle L. We know from the fluorescent pH titration of macrocycle L by acid/base that the change of fluorescence intersects at about pH=5.