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Abstract How consumer diversity determines consumption efficiency is a central issue in ecology. In the context of predation and biological control, this relationship concerns predator diversity and predation efficiency. Reduced predation efficiency can result from different predator taxa eating each other in addition to their common prey (interference due to intraguild predation). By contrast, multiple predator taxa with overlapping but complementary feeding niches can generate increased predation efficiency on their common prey (enemy complementarity). When viewed strictly from an ecological perspective, intraguild predation and enemy complementarity are opposing forces. However, from an evolutionary ecology perspective, predators facing strong intraguild predation may evolve traits that reduce their predation risk, possibly leading to niche complementarity between enemies; thus, selection from intraguild predation may lead to enemy complementarity rather than opposing it. As specialized predators that live in or on their hosts, parasitoids are subjected to intraguild predation from generalist predators that consume the parasitoids' hosts. The degree to which parasitoid–predator interactions are ruled by interference versus enemy complementarity has been debated. Here, we address this issue with field experiments in a forest community consisting of multiple species of trees, herbivorous caterpillars, parasitoids, ants, and birds. Our experiments and analyses found no interference effects, but revealed clear evidence for complementarity between parasitoids and birds (not ants). Parasitism rates by hymenopterans and dipterans were negatively associated with bird predation risk, and the variation in the strength of this negative association suggests that this enemy complementarity was due to parasitoid avoidance of intraguild predation. We further argue that avoidance of intraguild predation by parasitoids and other arthropod predators may explain enigmatic patterns in vertebrate–arthropod–plant food webs in a variety of terrestrial ecosystems. 

Quantitative labeling of biomolecules is necessary to advance areas of antibody–drug conjugation, super-resolution microscopy imaging of molecules in live cells, and determination of the stoichiometry of protein complexes. Bio-orthogonal labeling to genetically encodable noncanonical amino acids (ncAAs) offers an elegant solution; however, their suboptimal reactivity and stability hinder the utility of this method. Previously, we showed that encoding stable 1,2,4,5-tetrazine (Tet)-containing ncAAs enables rapid, complete conjugation, yet some expression conditions greatly limited the quantitative reactivity of the Tet-protein. Here, we demonstrate that reduction of on-protein Tet ncAAs impacts their reactivity, while the leading cause of the unreactive protein is near-cognate suppression (NCS) of UAG codons by endogenous aminoacylated tRNAs. To overcome incomplete conjugation due to NCS, we developed a more catalytically efficient tRNA synthetase and developed a series of new machinery plasmids harboring the aminoacyl tRNA synthetase/tRNA pair (aaRS/tRNA pair). These plasmids enable robust production of homogeneously reactive Tet-protein in truncation-free cell lines, eliminating the contamination caused by NCS and protein truncation. Furthermore, these plasmid systems utilize orthogonal synthetic origins, which render these machinery vectors compatible with any common expression system. Through developing these new machinery plasmids, we established that the aaRS/tRNA pair plasmid copy-number greatly affects the yields and quality of the protein produced. We then produced quantitatively reactive soluble Tet-Fabs, demonstrating the utility of this system for rapid, homogeneous conjugations of biomedically relevant proteins. 

An approach to varying the topology of nanobody assembly holds promise for creating more potent therapeutics and tools. 

Abstract The development of bioorthogonal fluorogenic probes constitutes a vital force to advance life sciences. Tetrazine‐encoded green fluorescent proteins (GFPs) show high bioorthogonal reaction rate and genetic encodability but suffer from low fluorogenicity. Here, we unveil the real‐time fluorescence mechanisms by investigating two site‐specific tetrazine‐modified superfolder GFPs via ultrafast spectroscopy and theoretical calculations. Förster resonance energy transfer is quantitatively modeled and revealed to govern the fluorescence quenching; for GFP150‐Tet with a fluorescence turn‐on ratio of ∼9, it contains trimodal subpopulations with good (P₁), random (P₂), and poor (P₃) alignments between the transition dipole moments of protein chromophore (donor) and tetrazine tag (Tet‐v2.0, acceptor). By rationally designing a more free/tight environment, we created new mutants Y200A/S202Y to introduce more P₂/P₁ populations and improve the turn‐on ratios to ∼14/31, making the fluorogenicity of GFP150‐Tet‐S202Y the highest among all up‐to‐date tetrazine‐encoded GFPs. In live eukaryotic cells, the GFP150‐Tet‐v3.0‐S202Y mutant demonstrates notably increased fluorogenicity, substantiating our generalizable design strategy. Key pointsUltrafast spectroscopy reveals FRET in action and inhomogeneous populations with different transition dipole moment alignments.Rational protein design of two new superfolder GFP mutants with record‐high fluorogenicity.Bioimaging application of the designed bioorthogonal protein mutant in live eukaryotic cells. 

Industrial emissions play a major role in the global methane budget. The Permian basin is thought to be responsible for almost half of the methane emissions from all U.S. oil- and gas-producing regions, but little is known about individual contributors, a prerequisite for mitigation. We use a new class of satellite measurements acquired during several days in 2019 and 2020 to perform the first regional-scale and high-resolution survey of methane sources in the Permian. We find an unexpectedly large number of extreme point sources (37 plumes with emission rates >500 kg hour −1 ), which account for a range between 31 and 53% of the estimated emissions in the sampled area. Our analysis reveals that new facilities are major emitters in the area, often due to inefficient flaring operations (20% of detections). These results put current practices into question and are relevant to guide emission reduction efforts.