Depsipeptide substrates for sortase-mediated N-terminal protein ligation

Technologies that allow the efficient chemical modification of proteins under mild conditions are widely sought after. Sortase-mediated peptide ligation provides a strategy for modifying the N or C terminus of proteins. This protocol describes the use of depsipeptide substrates (containing an ester linkage) with sortase A (SrtA) to completely modify proteins carrying a single N-terminal glycine residue under mild conditions in 4–6 h. The SrtA-mediated ligation reaction is reversible, so most labeling protocols that use this enzyme require a large excess of both substrate and sortase to produce high yields of ligation product. In contrast, switching to depsipeptide substrates effectively renders the reaction irreversible, allowing complete labeling of proteins with a small excess of substrate and catalytic quantities of sortase. Herein we describe the synthesis of depsipeptide substrates that contain an ester linkage between a threonine and glycolic acid residue and an N-terminal FITC fluorophore appended via a thiourea linkage. The synthesis of the depsipeptide substrate typically takes 2–3 d

Dual-targeting anti-angiogenic cyclic peptides as potential drug leads for cancer therapy

Peptide analogues derived from bioactive hormones such as somatostatin or certain growth factors have great potential as angiogenesis inhibitors for cancer applications. In an attempt to combat emerging drug resistance many FDA-approved anti-angiogenesis therapies are co-administered with cytotoxic drugs as a combination therapy to target multiple signaling pathways of cancers. However, cancer therapies often encounter limiting factors such as high toxicities and side effects. Here, we combined two anti-angiogenic epitopes that act on different pathways of angiogenesis into a single non-toxic cyclic peptide framework, namely MCoTI-II (Momordica cochinchinensis trypsin inhibitor-II), and subsequently assessed the anti-angiogenic activity of the novel compound. We hypothesized that the combination of these two epitopes would elicit a synergistic effect by targeting different angiogenesis pathways and result in improved potency, compared to that of a single epitope. This novel approach has resulted in the development of a potent, non-toxic, stable and cyclic analogue with nanomolar potency inhibition in in vitro endothelial cell migration and in vivo chorioallantoic membrane angiogenesis assays. This is the first report to use the MCoTI-II framework to develop a 2-in-1 anti-angiogenic peptide, which has the potential to be used as a form of combination therapy for targeting a wide range of cancers

Evolutionary social and biogeophysical changes in the Amazon, Ganges–Brahmaputra–Meghna and Mekong deltas

Policy-making in social-ecological systems increasingly looks to iterative, evolutionary approaches that can address the inherent complexity of interactions between human wellbeing, provision of goods, and the maintenance of ecosystem services. Here, we show how the analysis of available time-series in tropical delta regions over past decades can provide important insight into the social-ecological system dynamics in deltaic regions. The paper provides an exploratory analysis of the recent changes that have occurred in the major elements of three tropical deltaic social-ecological systems, such as demography, economy, health, climate, food, and water. Time-series data from official statistics, monitoring programmes, and Earth observation data are analysed to explore possible trends, slow and fast variables, and observed drivers of change in the Amazon, Ganges–Brahmaputra–Meghna and Mekong deltas. In the Ganges–Brahmaputra–Meghna delta zone, increasing gross domestic product and per capita income levels since the 1980s mirror rising levels of food and inland fish production. In contrast, non-food ecosystem services, such as water availability, water quality, and land stability appear to be deteriorating. In the Amazon delta, natural and anthropogenic perturbations are continuously degrading key ecosystem services, such as carbon storage in biomass and soils, the regulation of water balance, and the modulation of regional climate patterns. In the Mekong delta, rapid economic development, changing land-use practices, and salinity intrusion are progressively putting more pressure on the delivery of important provisioning services, such as rice and inland aquaculture production, which are key sources of staple food, farm incomes, and export revenue. Observed changes in many key indicators of ecosystem services point to a changing dynamic state and increased probability of systemic threshold transformations in the near future