NMR uncovers direct interaction between human NEDD4-1 and p34.

PTEN, an important tumor suppressor and a key regulator of the PI3K/AKT signaling pathway, is often deleted/mutated in different types of cancer. The E3 ubiquitin ligase NEDD4-1 catalyzes the polyubiquitination of PTEN, thereby acting as a negative regulator of PTEN. Stability of NEDD4-1, in turn, is tightly controlled by a 34 kDa oncoprotein, p34 and it regulates PTEN degradation and activates PI3K/AKT pathway, resulting in cancer metastasis.

Misassigned natural products and their revised structures.

Natural products are a major pipeline for drug development and are responsible for more than 50 % of drugs on the market. NMR is a fundamental and powerful tool for the structure determination of natural products. It is essential to provide unambiguous chemical structure information on natural products in drug development research, including the structure-activity relationship, derivatization and pharmacokinetic/pharmacodynamic studies.

CD-200 induces apoptosis and inhibits Bcr-Abl signaling in imatinib-resistant chronic myeloid leukemia with T315I mutation.

Chronic myeloid leukemia (CML) is characterized by a constitutively active Bcr-Abl tyrosine kinase. Although Imatinib has been proven to be an effective drug against CML, its resistance has been observed with disease relapse due to T315I predominant point mutation. Liriodendron tulipifera L.

Plant cell wall polysaccharides as potential resources for the development of novel prebiotics.

Prebiotic oligosaccharides, with a degree of polymerization (DP) of mostly less than 10, exhibit diverse biological activities that contribute to human health. Currently available prebiotics are mostly derived from disaccharides and simple polysaccharides found in plants. Subtle differences in the structures of oligosaccharides can cause significant differences in their prebiotic proper-ties.

In planta biocatalysis screen of P450s identifies 8-methoxypsoralen as a substrate for the CYP82C subfamily, yielding original chemical structures.

An in vivo plant screen that allows for the analysis of exogenously applied substrates against transgenic Arabidopsis lines overexpressing individual cytochrome P450s has been developed. By deploying this screen with a subset of 91 P450s, we have identified an original substrate for members of the CYP82C subfamily. The therapeutic compound 8-methoxypsoralen was hydroxylated by plants overexpressing CYP82C2 or CYP82C4, forming 5-hydroxy-8-methoxypsoralen.

Enhancement of alkaloid production in opium and California poppy by transactivation using heterologous regulatory factors.

Genes encoding regulatory factors isolated from Arabidopsis, soybean and corn have been screened to identify those that modulate the expression of genes encoding for enzymes involved in the biosynthesis of morphinan alkaloids in opium poppy (Papaver somniferum) and benzophenanthridine alkaloids in California poppy (Eschscholzia californica). In opium poppy, the over-expression of selected regulatory factors increased the levels of PsCOR (codeinone reductase), Ps4'OMT (S-adenosyl-l-methionine:3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase) and Ps6OMT [(R,S)-norcoclaurine 6-O-methyltransferase] transcripts by 10- to more than 100-fold. These transcriptional activations translated into an enhancement of alkaloid production in opium poppy of up to at least 10-fold.

Anti-HCV bioactivity of pseudoguaianolides from Parthenium hispitum.

Five new (1-5) and four known (6-9) C14-oxygenated 1alpha-hydroxy-11(13)-pseudoguaien-6beta,12-olides with potent inhibition of hepatitis C virus (HCV) replication were obtained from Parthenium hispitum via high-throughput natural product chemistry methods. A semipreparative HPLC system was used to purify these compounds. The miniaturization of the structure elucidation and dereplication for the mass-limited samples were performed primarily utilizing a capillary-scale NMR probe.

Bacterial biofilm inhibitors from Diospyros dendo.

One new (1) and four known (2-5) ursene triterpenes with potent inhibition of the formation of the bacterial biofilm Pseudomonas aeruginosa PA01 were obtained from Diospyros dendo using a high-throughput natural products chemistry procedure. These compounds were isolated as mass-limited samples. The miniaturization of the structure elucidation and dereplication was performed primarily utilizing a capillary-scale NMR probe.

Cyclolignans from Scyphocephalium ochocoa via high-throughput natural product chemistry methods.

Two 2,7'-cyclolignans, ocholignans A and B, were obtained as mass-limited samples from Scyphocephalium ochocoa via high-throughput natural products chemistry methods. The rapid structure elucidation of each compound was primarily facilitated by NMR data acquisition using a capillary-scale NMR probe, CapNMR probe. Ocholignan A was found to possess significant in vitro antibacterial activity against Gram-positive bacteria methicillin-resistant Staphylococcus aureus ATCC 33591 and S.

Application of capillary-scale NMR for the structure determination of phytochemicals.

Employing a capillary-scale NMR probe enables the miniaturisation of structure determination and de-replication of purified natural products from plants using only 5-100 microg of material. Approximately 5 microg are required to perform one-dimensional proton and two-dimensional homonuclear (COSY and NOESY) NMR experiments; some 30 microg are needed to acquire HMQC- or HSQC-NMR spectra; ca. 75-100 microg are necessary to measure HMBC-NMR spectra; and around 200 microg of a compound are needed to perform 13C- and DEPT-NMR experiments.

Miniaturization of the structure elucidation of novel natural products--two trace antibacterial acylated caprylic alcohol glycosides from Arctostaphylos pumila.

High-throughput isolation, purification and analysis methods applied to natural products libraries from plants gave rise to the discovery of two novel acylated caprylic alcohol glycosides (1, 2) produced by Arctostaphylos pumila. The NMR spectra were acquired using the CapNMR probe and performed on mass-limited samples, which enabled us to elucidate the structures of 2,6-diacetyl-3,4-diisobutyl-1- O-octylglucopyranoside (1, 200 microg) and 2,6-diacetyl-3,4-dimethylbutyl-1- O-octylglucopyranosid (2, 70 microg). Compounds 1 and 2 exhibited antibacterial activity against Gram-positive methicillin-resistant Staphylococcus aureus with an MIC of 128 microg/mL and 64 microg/mL, respectively.

Suaveolindole, a new mass-limited antibacterial indolosesquiterpene from Greenwayodendron suaveolens obtained via high-throughput natural products chemistry methods.

Utilizing high-throughput isolation, purification, and analysis methods applied to a natural products library, a new mass-limited antibacterial indolosesquiterpene, suaveolindole (1), was obtained from Greenwayodendron suaveolens. The miniaturization of the structure elucidation of 1 was performed primarily using the CapNMR probe. Compound 1 was found to possess significant in vitro antibacterial activity against the Gram-positive bacteria Bacillus subtilis (ATCC 43223), Staphylococcus aureus (ATTC 6538P), and methicillin-resistant Staphylococcus aureus (ATTC 33591), with MIC values of 4, 8, and 8 microg/mL, respectively.

A phthalide with in vitro growth inhibitory activity from an oidiodendron strain.

A screening campaign was implemented utilizing capillary electrophoresis as a primary assay to discover binders to the cancer target Akt1 from a crude natural extract library. Fungal extracts with binding activities were characterized for biochemical inhibition of Akt1 to phosphorylate the downstream substrate protein Bad. One of the crude extracts with bioactivity selected for isolation and structure elucidation from fermentation of the fungal culture Oidiodendron sp.

Identification of an endogenous ligand that activates pregnane X receptor-mediated sterol clearance.

The nuclear receptor PXR (pregnane X receptor) is a broad-specificity sensor that recognizes a wide variety of synthetic drugs and xenobiotic agents. On activation by these compounds, PXR coordinately induces a network of transporters, cytochrome P450 enzymes, and other genes that effectively clear xenobiotics from the liver and intestine. Like PXR, the majority of its target genes also possess a broad specificity for exogenous compounds.

Vidalenolone, a novel phenolic metabolite from the tropical red alga Vidalia sp.

The Indonesian red alga Vidalia sp. was identified as a candidate for fractionation because its crude lipid extract showed activity in a mechanism-based anticancer assay (Fyn SH2-inhibitory activity). A chemically novel phenolic metabolite, vidalenolone, as well as two previously described and structurally simple phenols, were isolated as SH2-inactive substances.

Development of a solid-phase binding assay and identification of nonpeptide ligands for the FynB Src homology 2 domain.

The nonreceptor tyrosine kinase FynB is known to be required in the induction of long-term potentiation (LTP), a cellular mechanism for learning and memory. Ligands of the FynB SH2 domain as a possible FynB activator are, thus, of great interest. In this study, a solid-phase ligand binding assay was established to meet the screening requirement of high-throughput and ease of use, and in an attempt to find the specific ligands for the FynB SH2 domain.