Plasmin activates epithelial Na+ channels by cleaving the gamma subunit

Proteolytic processing of epithelial sodium channel (ENaC) subunits occurs as channels mature within the biosynthetic pathway. The proteolytic processing events of the alpha and gamma subunits are associated with channel activation. Furin cleaves the alpha subunit ectodomain at two sites, releasing an inhibitory tract and activating the channel. However, furin cleaves the gamma subunit ectodomain only once. A second distal cleavage in the gamma subunit induced by other proteases, such as prostasin and elastase, is required to release a second inhibitory tract and further activate the channel. We found that the serine protease plasmin activates ENaC in association with inducing cleavage of the gamma subunit at gammaLys194, a site distal to the furin site. A gammaK194A mutant prevented both plasmin-dependent activation of ENaC and plasmin-dependent production of a unique 70-kDa carboxyl-terminal gamma subunit cleavage fragment. Plasmin-dependent cleavage and activation of ENaC may have a role in extracellular volume expansion in human disorders associated with proteinuria, as filtered plasminogen may be processed by urokinase, released from renal tubular epithelium, to generate active plasmin.     

Glycan Evolution in Response to Collaboration,Conflict, and Constraint

Glycans, oligo- and polysaccharides secreted or attached to proteins and lipids, cover the surfaces of all cells and have a regulatory capacity and structural diversity beyond any other class of biological molecule. Glycans may have evolved these properties because they mediate cellular interactions and often face pressure to evolve new functions rapidly. We approach this idea two ways. First, we discuss evolutionary innovation. Glycan synthesis, regulation, and mode of chemical interaction influence the spectrum of new forms presented to evolution. Second, we describe the evolutionary conflicts that arise when alleles and individuals interact. Glycan regulation and diversity are integral to these biological negotiations. Glycans are tasked with such an amazing diversity of functions that no study of cellular interaction can begin without considering them. We propose that glycans predominate the cell surface because their physical and chemical properties allow the rapid innovation required of molecules on the frontlines of evolutionary conflict.     

Scaffold oriented synthesis. Part 4: design, synthesis and biological evaluation of novel 5-substituted indazoles as potent and selective kinase inhibitors employing heterocycle forming and multicomponent reactions

We report the synthesis and biological evaluation of 5-substituted indazoles as kinase inhibitors. The compounds were synthesized in a parallel synthesis fashion from readily available starting materials employing heterocycle forming and multicomponent reactions and were evaluated against a panel of kinase assays. Potent inhibitors were identified for Gsk3β, Rock2, and Egfr.     

Synthesis and structure–activity relationship of piperidine-derived non-urea soluble epoxide hydrolase inhibitors

A series of potent amide non-urea inhibitors of soluble epoxide hydrolase (sEH) is disclosed. The inhibition of soluble epoxide hydrolase leads to elevated levels of epoxyeicosatrienoic acids (EETs), and thus inhibitors of sEH represent one of a novel approach to the development of vasodilatory and anti-inflammatory drugs. Structure–activities studies guided optimization of a lead compound, identified through high-throughput screening, gave rise to sub-nanomolar inhibitors of human sEH with stability in human liver microsomal assay suitable for preclinical development.     

A TEST OF THE CONJECTURE THAT G‐MATRICES ARE MORE STABLE THAN B‐MATRICES

The G‐matrix occupies an important position in evolutionary biology both as a summary of the inheritance of quantitative traits and as an ingredient in predicting how those traits will respond to selection and drift. Consequently, the stability of G has an important bearing on the accuracy of predicted evolutionary trajectories. Furthermore, G should evolve in response to stable features of the adaptive landscape and their trajectories through time. Although the stability and evolution of G might be predicted from knowledge of selection in natural populations, most empirical comparisons of G‐matrices have been made in the absence of such a priori predictions. We present a theoretical argument that within‐sex G‐matrices should be more stable than between‐sex B‐matrices because they are more powerfully exposed to multivariate stabilizing selection. We tested this conjecture by comparing estimates of B‐ and within‐sex G‐matrices among three populations of the garter snake Thamnophis elegans. Matrix comparisons using Flury's hierarchical approach revealed that within‐sex G‐matrices had four principal components in common (full CPC), whereas B‐matrices had only a single principal component in common and eigenvalues that were more variable among populations. These results suggest that within‐sex G is more stable than B , as predicted by our theoretical argument.     

Environment-specific heterozygote deficiency and developmental instability in hybrid Mytilus

The multiple discrete hybrid zones that characterize Mytilus blue mussels allow a novel, non-manipulative, examination of the selective pressures that create and maintain species. If endogenous genetic incompatibility is solely responsible for post-zygotic isolation, then individuals of a specified hybrid genotype are expected to show similar average fitness across environments. However, if hybrid fitness differs across environments, then exogenous selection is implicated, either via ecological selection or environment-specific expression of intrinsic genetic incompatibilities. Correspondence between developmental instability of hybrids and heterozygote deficiency, estimated in two M. trossulus×M. galloprovincialis hybrid zones on the coast of North America, indicates that environment-dependent selection against hybrids may contribute to reproductive isolation among Pacific Mytilus species.     

HIERARCHICAL COMPARISON OF GENETIC VARIANCE-COVARIANCE MATRICES. II COASTAL-INLAND DIVERGENCE IN THE GARTER SNAKE,THAMNOPHIS ELEGANS

The time-scale for the evolution of additive genetic variance-covariance matrices (G-matrices) is a crucial issue in evolutionary biology. If the evolution of G-matrices is slow enough, we can use standard multivariate equations to model drift and selection response on evolutionary time scales. We compared the G-matrices for meristic traits in two populations of gaiter snakes (Thamnophis elegans) with an apparent separation time of 2 million years. Despite considerable divergence in the meristic traits, foraging habits, and diet, these populations show conservation of structure in their G-matrices. Using Flury's hierarchial approach to matrix comparisons, we found that the populations have retained the principal components (eigenvectors) of their G-matrices, but their eigenvalues have diverged. In contrast, we were unable to reject the hypothesis of equal environmental matrices (E-matrices) for these populations. We propose that a conserved pattern of multivariate stabilizing selection may have contributed to conservation of G- and E-matrix structure during the divergence of these populations.