The structure and catalytic mechanism of a poly(ADP-ribose) glycohydrolase

Post-translational modification of proteins by poly(ADP-ribosyl)ation regulates many cellular pathways that are critical for genome stability, including DNA repair, chromatin structure, mitosis and apoptosis1. Poly(ADP-ribose) (PAR) is composed of repeating ADP-ribose units linked via a unique glycosidic ribose–ribose bond, and is synthesized from NAD by PAR polymerases1, 2. PAR glycohydrolase (PARG) is the only protein capable of specific hydrolysis of the ribose–ribose bonds present in PAR chains; its deficiency leads to cell death3, 4. Here we show that filamentous fungi and a number of bacteria possess a divergent form of PARG that has all the main characteristics of the human PARG enzyme. We present the first PARG crystal structure (derived from the bacterium Thermomonospora curvata), which reveals that the PARG catalytic domain is a distant member of the ubiquitous ADP-ribose-binding macrodomain family5, 6. High-resolution structures of T. curvata PARG in complexes with ADP-ribose and the PARG inhibitor ADP-HPD, complemented by biochemical studies, allow us to propose a model for PAR binding and catalysis by PARG. The insights into the PARG structure and catalytic mechanism should greatly improve our understanding of how PARG activity controls reversible protein poly(ADP-ribosyl)ation and potentially of how the defects in this regulation are linked to human disease

Real-time monitor on the release of salicylic acid from chitosan gel beads by means of dielectric spectroscopy

The release processes of salicylic acid (SA) from the chitosan gel beads (CGB) with different crosslinking densities were monitored in real time by means of dielectric spectroscopy. Distinct dielectric relaxations attributed to interfacial polarization were observed, which exhibited obvious dependences on the crosslinking density and the release time. In line with Hanai's equation, the properties of CGB and release media were quantitatively determined from the dielectric profiles. Based on these properties and dielectric behaviors, the release profiles were analyzed. It was found that only the dispersion of CGB with the lowest crosslinking density exhibited release behavior, while other dispersions showed an obvious adsorption process following a burst of drug diffusion. The total loading and release amount of SA were determined and compared, and the influence of crosslinking degree on the drug loading and release was discussed. In addition, the release behavior was confirmed to be diffusionally controlled, and the diffusion coefficient of SA was determined