Water-Mediated Carbon–Oxygen Hydrogen Bonding Facilitates S-Adenosylmethionine Recognition in the Reactivation Domain of Cobalamin-Dependent Methionine Synthase

The C-terminal domain of cobalamin-dependent methionine synthase (MetH) has an essential role in catalyzing the reactivation of the enzyme following the oxidation of its cobalamin cofactor. This reactivation occurs through reductive methylation of the cobalamin using S-adenosylmethionine (AdoMet) as the methyl donor. Herein, we examine the molecular recognition of AdoMet by the MetH reactivation domain utilizing structural, biochemical, and computational approaches. Crystal structures of the Escherichia coli MetH reactivation domain in complex with AdoMet, the methyl transfer product S-adenosylhomocysteine (AdoHcy), and the AdoMet analogue inhibitor sinefungin illustrate that the ligands exhibit an analogous conformation within the solvent-exposed substrate binding cleft of the enzyme. AdoMet binding is stabilized by an intramolecular sulfur–oxygen chalcogen bond between the sulfonium and carboxylate groups of the substrate and by water-mediated carbon–oxygen hydrogen bonding between the sulfonium cation and the side chains of Glu1097 and Glu1128 that bracket the substrate binding cleft. AdoMet and sinefungin exhibited similar binding affinities for the MetH reactivation domain, whereas AdoHcy displayed an affinity for the enzyme that was an order of magnitude lower. Mutations of Glu1097 and Glu1128 diminished the AdoMet/AdoHcy binding selectivity ratio to approximately 2-fold, underscoring the role of these residues in enabling the enzyme to discriminate between the substrate and product. Together, these findings indicate that Glu1097 and Glu1128 in MetH promote high-affinity recognition of AdoMet and that sinefungin and potentially other AdoMet-based methyltransferase inhibitors can abrogate MetH reactivation, which would result in off-target effects associated with alterations in methionine homeostasis and one-carbon metabolism

Structural and Functional Characterization of Sulfonium Carbon-Oxygen Hydrogen Bonding in the Deoxyamino Sugar Methyltransferase TyIM1

The N-methyltransferase TylM1 from Streptomyces fradiae catalyzes the final step in the biosynthesis of the deoxyamino sugar mycaminose, a substituent of the antibiotic tylosin. The high-resolution crystal structure of TylM1 bound to the methyl donor S-adenosylmethionine (AdoMet) illustrates a network of carbon-oxygen (CH•••O) hydrogen bonds between the substrate’s sulfonium cation and residues within the active site. These interactions include hydrogen bonds between the methyl and methylene groups of the AdoMet sulfonium cation and the hydroxyl groups of Tyr14 and Ser120 in the enzyme. To examine the functions of these interactions, we generated Tyr14 to phenylalanine (Y14F) and Ser120 to alanine (S120A)mutations to selectively ablate the CH•••O hydrogen bonding to AdoMet. The TylM1 S120A mutant exhibited a modest decrease in the catalytic efficiency relative to wild type (WT) enzyme, whereas the Y14F mutation resulted in an approximately 30-fold decrease in catalytic efficiency. In contrast, site-specific substitution of Tyr14 by the noncanonical amino acid p-aminophenylalanine partially restored activity comparable to the WT enzyme. Correlatively, quantum mechanical calculations of the activation barrier energies of WT TylM1 and the Tyr14 mutants suggest that substitutions which abrogate hydrogen bonding with the AdoMet methyl group impair methyl transfer. Together, these results offer insights into roles of CH•••O hydrogen bonding in modulating the catalytic efficiency of TylM1

Linking global drivers of agricultural trade to on-the-ground impacts on biodiversity.

Consumption of globally traded agricultural commodities like soy and palm oil is one of the primary causes of deforestation and biodiversity loss in some of the world's most species-rich ecosystems. However, the complexity of global supply chains has confounded efforts to reduce impacts. Companies and governments with sustainability commitments struggle to understand their own sourcing patterns, while the activities of more unscrupulous actors are conveniently masked by the opacity of global trade. We combine state-of-the-art material flow, economic trade, and biodiversity impact models to produce an innovative approach for understanding the impacts of trade on biodiversity loss and the roles of remote markets and actors. We do this for the production of soy in the Brazilian Cerrado, home to more than 5% of the world´s species. Distinct sourcing patterns of consumer countries and trading companies result in substantially different impacts on endemic species. Connections between individual buyers and specific hot spots explain the disproportionate impacts of some actors on endemic species and individual threatened species, such as the particular impact of European Union consumers on the recent habitat losses for the iconic giant anteater (Myrmecophaga tridactyla). In making these linkages explicit, our approach enables commodity buyers and investors to target their efforts much more closely to improve the sustainability of their supply chains in their sourcing regions while also transforming our ability to monitor the impact of such commitments over time.UK Global Food Security programme (Project 304 BB/N02060X/1

Growing your own

A publication containing advice on a wide range of gardening topics, including composting, container gardens, fall/winter gardens, fertilizing, insect pests, plant diseases, planting guidelines, raised beds, site selection, slugs, soil improvement, tilling, warm-season crops, watering, and weeds. Includes regional tips for various parts of Oregon.Published April 2011. Replaces GROW. Facts and recommendations in this publication may no longer be valid. Please look for up-to-date information in the OSU Extension Catalog: http://extension.oregonstate.edu/catalogKeywords: soil preparation, gardening, pest control, Oregon, vegetable

A semi-continuous formulation for goal-oriented reduced-order models: 1D problems

A semi-continuous formulation is introduced for finding bases that minimise the error in a specific output functional of a reduced-order model. The formulation is advantageous in that it can be used with arbitrary reference data and can be easily applied to nonlinear models and functionals. A general description of the approach is given; then, explicit formulations are derived for the advection-diffusion and Burgers' equations. Numerical results are given for both linear and nonlinear functionals. These show substantial reductions in error when compared with POD modes, depending on the functional considered. The optimisation of bases for a reduced-order model using an approximated governing equation is also described, for which large increases in accuracy are obtained relative to POD modes. Copyright (C) 2015 John Wiley & Sons, Ltd

LONGBOW Supersonic Interceptor

A recent white paper entitled "From the Sea" has spotlighted the need for Naval Aviation to provide overland support to joint operations. The base for this support, the Aircraft Carrier (CVN), will frequently be unable to operate within close range of the battleground because of littoral land-based air and subsurface threats. A high speed, long range, carrier capable aircraft would allow the CVN to provide timely support to distant battleground operations. Such an aircraft, operating as a Deck-Launched Interceptor (DLI), would also be an excellent counter to Next Generation Russian Naval Aviation (NGRNA) threats consisting of supersonic bombers, such as the Backfire, equipped with the next generation of high-speed, long-range missiles. Additionally, it would serve as an excellent high speed Reconnaissance airplane, capable of providing Battle Force commanders with timely, accurate pre-mission targeting information and post-mission Bomb Damage Assessment (BDA). Recent advances in computational hypersonic airflow modeling has produced a method of defining aircraft shapes that fit a conical shock flow model to maximize the efficiency of the vehicle. This "Waverider" concept provides one means of achieving long ranges at high speeds. A Request for Proposal (RFP) was issued by Professor Conrad Newberry that contained design requirements for an aircraft to accomplish the above stated missions, utilizing Waverider technology. The FASTGUYS Corporation's response to this RFP is a design called the LONGBOW. Table I outlines the significant detailed requirements and compares LONGBOW'S performance with them.Naval Postgraduate School, Monterey, California.Approved for public release; distribution is unlimited