Radical S-adenosylmethionine (SAM) enzymes in cofactor biosynthesis: a treasure trove of complex organic radical rearrangement reactions.

In this minireview, we describe the radical S-adenosylmethionine enzymes involved in the biosynthesis of thiamin, menaquinone, molybdopterin, coenzyme F420, and heme. Our focus is on the remarkably complex organic rearrangements involved, many of which have no precedent in organic or biological chemistry.

Polyamide recognition-mass spectrometry for distinguishing hairpin DNA from coil DNA.

The discrimination between hairpin DNA and coil DNA has been well achieved through polyamides as probes by electrospray ionization (ESI) mass spectrometry. ESI mass spectra showed that polyamides bind to hairpin DNA with high selectivity, and almost no binding with coil DNA. In addition, the noncovalent interaction between polyamides and hairpin DNA was also studied; the results show that hairpin DNA with longer stem and polyamides with more heterocycles have higher binding affinity and stability in gas phase.

Spectroscopy probing of the formation, recognition, and conversion of a G-quadruplex in the promoter region of the bcl-2 oncogene.

This study has demonstrated the formation of the G-quadruplex structure from the G-rich sequence in the promoter region of the bcl-2 oncogene; the formation could be induced by addition of NH(4)(+) or K(+) ions. The binding affinity and stoichiometry of seven small molecules with the G-quadruplex were examined by using ESI-MS, as well as CD and UV spectroscopy. The binding-affinity order was determined to be P1 approximately = P5 > P2 > P3 approximately = P4 > P7 > P6. In particular, the small-molecule induction of the structural transition between the G-quadruplex and duplex DNA forms in this promoter region was investigated by ESI-MS. We directly observed specific binding of dehydrocorydaline (P7) and cationic porphyrin (P5) in one system consisting of the G-quadruplex and the duplex DNA, respectively. The results indicate that P7 selectively stabilizes the G-quadruplex and shifts the equilibrium toward G-quadruplex formation of the bcl-2 promoter, whereas P5 converts the G-quadruplex into the duplex DNA, which results in strong and selective binding to the duplex form. Therefore, P5 and P7 with their attractive binding specificities could be considered as precursors for pathway-specific drug design for regulation of bcl-2 oncogene transcription.

[Study on HPLC fingerprint of Trachelospermum jasminoides].

OBJECTIVE: To establish HPLC fingerprint for identification and evaluation of the quality of Trachelospermum jasminoides. METHOD: The analysis was performed on a ZORBAX Eclipse XDB-C18 analytical column (4.6 mm x 150 mm, 5 microm) with H2O (A) and methanol (B) as mobile phases in gradient mode. The elutin conditions were 0-15 min, changed from 10% B to 30% B; 15-40 min, to 40% B; 40-60 min, to 60% B. The column temperature was set at 30 degrees C and the flow rate was 0.8 mL x min(-1) with the detection wavelength of 230 nm. RESULT: The HPLC chromatographic fingerprint of T. jasminoides, showing 19 characteristic peaks, was established from 14 lots of T. jasminoides. The similarity of every lot of T. jasminoides was calculated as good in general, the similarity of 10 lots was above 0.9 and the other 4 lots had low similarity. CONCLUSION: The chromatographic fingerprint of T. jasminoides with high characteristics and specificity can be used to control its quality.

Controllable mullite bismuth ferrite micro/nanostructures with multifarious catalytic activities for switchable/hybrid catalytic degradation processes.

In this work, controllable preparation of micro/nanostructured bismuth ferrites (BFOs) were used to investigate multifarious heterogeneous catalyses, including Fenton/Fenton-like reaction, photocatalysis, photo-Fenton oxidation, and peroxymonosulfate (PMS) activation. Results showed that BFO can be used asa novel catalyst to activate switchable catalytic degradation of organic matters. Additionally, a novel catalytic system for degradation of organic pollutants, which integrating all-above heterogeneous catalyses is denoted as BFO/H2O2/PMS hybrid reaction, is introduced for the first time. BFO/H2O2/PMS system effectively degraded>99% for both methyl orange (MO) and sulfamethoxazole (SMX) within 60min, which shows better efficiency than above BFO-driven catalyses. The major SMX degradation pathway in BFO/H2O2/PMS system is proposed via detecting intermediates using LC/MS/MS. It was found that catalytic activities of BFOs are in the order of BFO-L (co-precipitation, micro/nanosize, single crystals exposing facet (001))>BFO-H (hydrothermal, nanocluster with a higher surface area than other BFOs)>BFO-C (fabricated using calcination process, microsize), which demonstrated that crystallographic orientation is more significant in heterogeneous catalyses than specific surface area at micro/nanoscale. Besides, the required H2O2 consumption for achieving 99% TOC removal was identified in BFO-driven photo-Fenton oxidation. The other effects on degradation efficiency, such as H2O2 dosage and pH, were investigated as well. In Fenton/Fenton-like reaction, reaction conditions suggested are ∼61.5mM H2O2 dosage and pH≥4.5 to avoid quenching of HO into HO2 by excessive H2O2 and Fe leaching.

Monitoring protein-small molecule interactions by local pH modulation.

We have developed a technique for sensing protein-small molecule and protein-ion interactions in bulk aqueous solution by utilizing a pH sensitive dye, 5-(and-6)-carboxyfluorescein, conjugated to free lysine residues on the surfaces of designated capture proteins. The fluorescein intensity was found to change by about 6% and 15% for small molecule and ion binding, respectively. The assay works by modulating the local electric fields around a pH sensitive dye. This, in turn, alters the dye's apparent pK(A) value. Such changes may result directly from the charge on the analyte, occur through allosteric effects related to the binding process, or result from a combination of both. The assay was used to follow the binding of Ca(2+) to calmodulin (CaM) and thiamine monophosphate (ThMP) to thiamine binding protein A (TbpA). The results demonstrate a binding constant of 1.1 µM for the Ca(2+)/CaM pair and 3.2 nM for ThMP/TbpA pair, which are in excellent agreement with literature values. These assays demonstrate the generality of this method for observing the interactions of small molecules and ions with capture proteins. In fact, the assay should work as a biosensor platform for most proteins containing a specific ligand binding site, which would be useful as a simple and rapid preliminary screen of protein-ligand interactions.

Discrimination of cyclic peptide diastereomers by electrospray ionization tandem mass spectrometry.

In this research, the characteristic ions' abundance ratio between two isomers A and B in MS/MS mass spectra was defined as a parameter for discriminating diastereomers. Through this ratio, the discrimination of four pairs of cyclic peptide (CP) diastereomers was successfully achieved. Furthermore, in the analysis of diastereomers' mixtures, both calibration curve and calculational methods were substantiated to have high precision and accuracy. The average absolute errors of the two methods were 2.0 and 2.5% in the 48 measurements of 16 samples, respectively. This research provided a promising approach for the analysis of the CP diastereomers in the fields of asymmetrical synthesis, chiral natural products and structural biology by ESI-MS/MS.