Mechanistic inferences from the binding of ligands to LpxC, a metal-dependent deacetylase

The metal-dependent deacetylase LpxC catalyzes the first committed step of lipid A biosynthesis in Gram-negative bacteria. Accordingly, LpxC is an attractive target for the development of inhibitors that may serve as potential new antibiotics for the treatment of Gram-negative bacterial infections. Here, we report the 2.7 A resolution X-ray crystal structure of LpxC complexed with the substrate analogue inhibitor TU-514 and the 2.0 A resolution structure of LpxC complexed with imidazole. The X-ray crystal structure of LpxC complexed with TU-514 allows for a detailed examination of the coordination geometry of the catalytic zinc ion and other enzyme-inhibitor interactions in the active site. The hydroxamate group of TU-514 forms a bidentate chelate complex with the zinc ion and makes hydrogen bond interactions with conserved active site residues E78, H265, and T191. The inhibitor C-4 hydroxyl group makes direct hydrogen bond interactions with E197 and H58. Finally, the C-3 myristate moiety of the inhibitor binds in the hydrophobic tunnel of the active site. These intermolecular interactions provide a foundation for understanding structural aspects of enzyme-substrate and enzyme-inhibitor affinity. Comparison of the TU-514 complex with cacodylate and imidazole complexes suggests a possible substrate diphosphate binding site and highlights residues that may stabilize the tetrahedral intermediate and its flanking transition states in catalysis. Evidence of a catalytic zinc ion in the native zinc enzyme coordinated by H79, H238, D242, and two water molecules with square pyramidal geometry is also presented. These results suggest that the native state of this metallohydrolase may contain a pentacoordinate zinc ion, which contrasts with the native states of archetypical zinc hydrolases such as thermolysin and carboxypeptidase A. © 2006 American Chemical Society.link_to_subscribed_fulltex

State of the art in the development and properties of protein-based films and coatings and their applicability to cellulose based products: an extensive review

There is increasing research towards the substitution of petrochemicals by sustainable components. Biopolymers such as proteins, polysaccharides, and lipids derive from a variety of crop sources and most promisingly from waste streams generated during their processing by the agro food industry. Among those, proteins of different types such as whey, casein, gelatin, wheat gluten, soy protein or zein present a potential beyond the food and feed industry for the application in packaging. The general protein hydrophilicity promotes a good compatibility to polar surfaces, such as paper, and a good barrier to apolar gases, such as oxygen and carbon dioxide. The present review deals with the development of protein-based coatings and films. It includes relevant discussion for application in paper or board products, as well as an outlook on its future industrial potential. Proteins with suitable functionalities as food packaging materials are described as well as the different technologies for processing the coatings and the current state of the art about the coating formulations for selectively modulating barrier, mechanical, surface and end of life properties. Some insights onto regulations about packaging use, end of life and perspectives of such natural coating for decreasing the environmental impact of packages are give