Intracellular directed evolution of proteins from combinatorial libraries based on conditional phage replication
Brodel AK, Jaramillo A, Isalan M
Directed evolution is a powerful tool to improve the characteristics of biomolecules. Here we present a protocol for the intracellular evolution of proteins with distinct differences and advantages in comparison with established techniques. These include the ability to select for a particular function from a library of protein variants inside cells, minimizing undesired coevolution and propagation of nonfunctional library members, as well as allowing positive and negative selection logics using basally active promoters. This protocol requires no special equipment, allowing synthetic biologists and general users to evolve improved biomolecules within ~7 weeks.
The mechanistic link between Arc/Arg3.1 expression and AMPA receptor endocytosis
Wall, M. (Mark) and Corrêa, Sônia A. L
The activity-regulated cytoskeleton associated protein (Arc/Arg3.1) plays a key role in determining synaptic strength through facilitation of AMPA receptor (AMPAR) endocytosis. Here we review data on the link between Arc expression and the clathrin-mediated endocytic pathway which internalises AMPARs and discuss the significance of Arc binding to the clathrin adaptor protein 2 (AP-2) and to endophilin/dynamin. We consider which AMPAR subunits are selected for Arc-mediated internalisation, implications for synaptic function and consider Arc as a therapeutic target.
Subcellular localisation and interactions among rubber particle proteins from Hevea braziliensis
We report the subcellular localization and interactions between selected isoforms of the major proteins of rubber particles from Hevea braziliensis. We show that these proteins associate with the endoplasmic reticulum membrane and can interact with the key cytosolic biosynthetic enzyme, cis-prenyltransferase. These data provide initial information on how the rubber transferase complex may assemble at the endoplasmic reticulum membrane. Natural rubber (polyisoprene) from the rubber tree Hevea braziliensis is synthesised by specialised cells called laticifers. It is not clear how rubber particles arise, although one hypothesis is that they derive from the endoplasmic reticulum (ER) membrane.
Pedal peptide/orcokinin-type neuropeptide signaling in a deuterostome: the anatomy and pharmacology of starfish myorelaxant peptide in Asterias rubens
Ming Lin, Michaela Egertova, Cleidiane G Zampronio, Alexandra M Jones, Maurice R Elphic
Pedal peptide (PP) and orcokinin (OK) are related neuropeptides that were discovered in protostomian invertebrates (mollusks, arthropods). However, analysis of genome/transcriptome sequence data has revealed that PP/OK-type neuropeptides also occur in a deuterostomian phylum—the echinoderms. Furthermore, a PP/OK-type neuropeptide (starfish myorelaxant peptide, SMP) was recently identified as a muscle relaxant in the starfish Patiria pectinifera. The results showed from mass spectometry showed that divergence of deuterostomes and protostomes may have been accompanied by an inhibitory–excitatory transition in the roles of PP/OK-type neuropeptides as regulators of muscle activity.
Distribution, Community Composition, and Potential Metabolic Activity of Bacterioplankton in an Urbanized Mediterranean Sea Coastal Zone
Kumari Richa, Cecilia Balestra, Roberta Piredda, Vladimir Benes, Marco Borra, Augusto Passarelli, Francesca Margiotta, Maria Saggiomo, Elio Biffali, Remo Sanges, David J. Scanlan, and Raffaella Casotti
Bacterioplankton are fundamental components of marine ecosystems and also influence the entire biosphere by contributing to the global biogeochemical cycles of key elements. Yet, there is a significant gap in knowledge about their diversity, specific activity and environmental factors that shape their community composition and function. Here, the distribution and diversity of surface bacterioplankton along the coastline of the Gulf of Naples was investigated.
A cheminformatics review of auxins as herbicides
Mussa Quareshy, Justyna Prusinska, Jun Li, Richard Napier
Herbicides are an important asset in ensuring food security, especially when faced with ever-increasing demand on food production to feed the planet. The current selection of herbicides is increasingly encountering resistance in the agricultural weeds they once targeted effectively. It is imperative that new compounds, or even better, new modes of actions are discovered in order to overcome such resistances. This cheminformatics review looks at the current herbicides and evaluates their physiochemical properties on a class by class basis. We introduce an interactive, open source cheminformatics tool, complete with records for over 300 herbicidal compounds, and hope this helps for not only auxin discovery, but agrochemical discovery generally.