Solvation study of the non-specific lipid transfer protein from wheat by intermolecular NOEs with water and small organic molecules

Intermolecular nuclear Overhauser effects (NOEs) were measured between the protons of various small solvent or gas molecules and the non-specific lipid transfer protein (ns-LTP) from wheat. Intermolecular NOEs were observed with the hydrophobic pocket in the interior of wheat ns-LTP, which grew in intensity in the order cyclopropane (saturated solution) < methane (140 bar) < ethane (40 bar) < acetonitrile (5% in water) < cyclohexane (saturated solution) < benzene (saturated solution). No intermolecular NOEs were observed with dioxane (5% in water). The intermolecular NOEs were negative for all of the organic molecules tested. Intermolecular NOEs between wheat ns-LTP and water were weak or could not be distinguished from exchange-relayed NOEs. As illustrated by the NOEs with cyclohexane versus dioxane, the hydrophobic pocket in wheat ns-LTP preferably binds non-polar molecules. Yet, polar molecules like acetonitrile can also be accommodated. The pressure dependence of the NOEs between methane and wheat ns-LTP indicated incomplete occupancy, even at 190 bar methane pressure. In general, NOE intensities increased with the size of the ligand molecule and its vapor pressure. NMR of the vapor phase showed excellent resolution between the signals from the gas phase and those from the liquid phase. The vapor concentration of cyclohexane was fivefold higher than that of the dioxane solution, supporting the binding of cyclohexane versus uptake of dioxane.     

A role for the Psi-U mismatch in the recognition of the 5' splice site of yeast introns by the U1 small nuclear ribonucleoprotein particle

The U1 small nuclear ribonucleoprotein particle (snRNP)/5' splice site (5'SS) interaction in yeast is essential for the splicing process and depends on the formation of a short RNA duplex between the 5' arm of U1 snRNA and the 1st intronic nucleotides. This RNA/RNA interaction is characterized by the presence of a mismatch that occurs with almost all yeast introns and concerns nucleotides 4 on the pre-mRNA (a U) and 5 on U1 snRNA (a Psi). The latter nucleotide is well conserved from yeast to vertebrates, but its role in yeast and the significance of the associated mismatch in the U1 snRNA/5'SS interaction have never been fully explained. We report here that the presence of this mismatch is a determinant of stability that mainly affects the off rate of the interaction. To our knowledge this is the first report assigning a function to this noncanonical interaction. We also performed SELEX (systematic evolution of ligands by exponential enrichment) experiments by immunoprecipitating U1 snRNP and the associated RNA. The artificial phylogeny derived from these experiments allows the isolation of the selective pressure due to U1 snRNP binding on the 5'SS of yeast introns.     

Functional SARS-CoV-2-Specific Immune Memory Persists after Mild COVID-19

1. Download : Download high-res image (133KB)
2. Download : Download full-size image

     

Recent trends and novel concepts in cofactor-dependent biotransformations

Cofactor-dependent enzymes catalyze a broad range of synthetically useful transformations. However, the cofactor requirement also poses economic and practical challenges for the application of these biocatalysts. For three decades, considerable research effort has been devoted to the development of reliable in situ regeneration methods for the most commonly employed cofactors, particularly NADH and NADPH. Today, researchers can choose from a plethora of options, and oxidoreductases are routinely employed even on industrial scale. Nevertheless, more efficient cofactor regeneration methods are still being developed, with the aim of achieving better atom economy, simpler reaction setups, and higher productivities. Besides, cofactor dependence has been recognized as an opportunity to confer novel reactivity upon enzymes by engineering their cofactors, and to couple (redox) biotransformations in multi-enzyme cascade systems. These novel concepts will help to further establish cofactor-dependent biotransformations as an attractive option for the synthesis of biologically active compounds, chiral building blocks, and bio-based platform molecules.     

Genetic analysis reveals the costs of peri-urban development for the endangered grassland earless dragon

Australia’s natural temperate grasslands have diminished to 0.5 % of their former area since European settlement and, as a consequence, are highly fragmented and modified. Many vertebrate species that live in temperate grasslands are habitat specialists and therefore are at risk of decline through habitat loss and fragmentation. The grassland earless dragon (Tympanocryptis pinguicolla) is one such species. Once widespread, T. pinguicolla is now restricted to two general locations; the first is near Canberra in the Australian Capital Territory (including some adjacent land near Queanbeyan), and the second is the Monaro Tablelands in New South Wales. Here, we use microsatellite DNA data collected from the largest remaining populations near Canberra to examine genetic structure in this species in the context of the rapidly expanding urban landscape in this region. Our study revealed that, despite separation by only relatively small distances (largest distance ~13 km), the T. pinguicolla populations are highly genetically structured with little admixture. Our analyses also revealed that the population with the largest census size, but which has recently crashed in population size, exhibited little detectable gene flow to other populations and is essentially isolated. Our data indicate that significant barriers to dispersal exist among the remaining T. pinguicolla populations and that management of this species cannot rely on natural dispersal to bolster declining populations. Many different agencies and landholders are responsible for the protection of these remnant populations and a co-ordinated effort is required to provide reasonable confidence that the species will persist.