A new NMR method to monitor solvent-localized NMR signals in the two-phase liquid system is suggested. This method based on intermolecular single-quantum coherence (iSQC). Here, we exploited the feature of the local action of distant dipolar field (DDF) effect in order to filter out specific NMR signals dissolved in different solvents. This solvent specific iSQC spectroscopy was carried out on a model two-phase liquid system (D-glucose in water/palmitic acid in chloroform), and showed solvent-localized NMR signals. We believe our approaches might be useful in metabolic analysis such as two-phase liquid extraction scheme for labile chemical species.

Fipronil, the phenylpyrazole insecticide, is effective and used in various fields. Especially, fipronil was reliable because it was known to be specific on invertebrate animals than vertebrate animals including mammals. However, fipronil had potential risks that affect vertebrate animals as it blocks the gamma-aminobutyric acid (GABA) receptors that also exists in vertebrates as well as invertebrates. Therefore, it was necessary that harmful effects of fipronil on vertebrates are clarified. For this purpose, the zebrafish (Danio rerio) were used on behalf of vertebrate animals in present study. The zebrafish were exposed to 5 ㎍/L, 25 ㎍/L, and 50 ㎍/L of fipronil during 12, 24 and 72 hours. To closely observe toxic process, 12 hours and 24 hours of additional time point were set in the exposure test. Nuclear magnetic resonance (NMR)-based metabolomics is an approach to detect metabolic changes in organism resulted from external stimuli. In this study, NMR-based metabolomics showed the metabolic changes in zebrafish caused by fipronil exposure. Metabolic analysis revealed that fipronil interfered with energy metabolism and decreased the antioxidant ability in zebrafish. Antioxidant ability decline was remarkable at high exposure concentration. In addition, metabolic analysis results over time suggested that reactions for alleviating the excessive nerve excitation occurred in zebrafish after fipronil exposure. Through this study, it was elucidated that the adverse effects of fipronil on vertebrate animals are evident. The risk of fipronil on vertebrates can be no longer ignored. Moreover, this study has a meaning of practically necessary research for organism by examining the effects of fipronil at low concentrations existed in real environment.

Transcription factors are proteins that bind specific sites or elements in regulatory regions of DNA, known as promoters or enhancers, where they control the transcription or expression of target genes. MEIS1 protein is a DNA-binding domain present in human transcription factors and plays important roles in various biological functions. The hydrogen exchange rate constants of the imino protons were determined for the wild-type containing the consensus DNA-binding site for the MEIS1 and those of the mutant DNA duplexes using NMR spectroscopy. The G2A-, A3G- and C4T-mutant DNA duplexes lead to clear changes in thermal stabilities of these four consensus base pairs. These unique dynamic features of the four base pairs in the consensus 5'-TGAC-3' sequence might play crucial roles in the effective DNA binding of the MEIS1 protein.

All plants in natural ecosystems are living in symbiosis with endophytes. Recently, there has been an increasing interest in endophytes since these organisms can interact with the hosts and produce various structurally or biologically interesting molecules. This study aimed to identify these molecules from endophytes. Chemical investigation of Climacium dendroides-derived fungus Talaromyces sp. resulted in the isolation of two diphenyl ether derivatives, purpactin A (1) and penicillide (2), and two steroids, dankasterone A (3) and calvasterol B (4). The structures of the compounds were identified via extensive spectroscopic and spectrometric methods. Four compounds did not show any antioxidative activities in the on-line antioxidant activity screening system.

In a relatively short period, monoclonal antibodies have made dramatic success as therapeutics for various diseases such as cancers and autoimmune diseases and become an important development items for many pharmaceutical companies. In order to develop antibody drug, it is important to investigate the structural characteristics of both antibody and antigen. NMR studies on antibody are extremely challenging due to big huddles such as a big size of protein and isotope labeling, nevertheless, several studies have been reported in 10 years. Here, we analyzed 95 papers dealing with antibody-related NMR studies reported in recent 10 years. We categorized papers into 3 types: 1) structural characterization of antibody, 2) structural characterization of antigen using antibody, 3) amyloidosis caused by fragment of antibody. This work would shed new light on antibody-related NMR studies.

Apolipoprotein B-100 (apo B-100), the main protein component that makes up LDL (Low density lipoprotein), consists of 4,536 amino acids and serves to combine with the LDL receptor. The oxidized LDL peptides by malondialdehyde (MDA) or acetylation in vivo were act as immunoglobulin (Ig) antigens and peptide groups were classified into 7 peptide groups with subsequent 20 amino acids (P1-P302). The biomimetic peptide P99 (KGTYG LSCQR DPNTG RLNGE) out of B-group peptides carrying the highest value of IgM antigens were selected for structural studies that may provide antigen specificity. Circular Dichroism (CD) spectra were measured for peptide secondary structure in the range of 190-260 nm. Experimental results show that P99 has pseudo α-helice and random coil structure. Homonuclear (COSY, TOCSY, NOESY) 2D-NMR experiments were carried out for NMR signal assignments and structure determination for P99. On the basis of these completely assigned NMR spectra and proton distance information, distance geometry (DG) and molecular dynamic (MD) were carried out to determine the structures of P99. The proposed structure was selected by comparisons between experimental NOE spectra and back-calculated 2D NOE results from determined structure showing acceptable agreement. The total Root-Mean-Square-Deviation (RMSD) value of P99 obtained upon superposition of all atoms were in the set range. The solution state P99 has mixed structure of pseudo α-helix and β-turn(Gln[9] to Thr[13]). These NMR results are well consistent with secondary structure from experimental results of circular dichroism. Structural studies based on NMR may contribute to the prevent oxidation studies of atherosclerosis and observed conformational characteristics of apo B-100 in LDL using monoclonal antibodies.