“Stoichioproteomics” relates the elemental composition of proteins and proteomes to variation in the physiological and ecological environment. To help harness and explore the wealth of hypotheses made possible under this framework, we introduce GRASP (http://www.graspdb.net), a public bioinformatic knowledgebase containing information on the frequencies of 20 amino acids and atomic composition of their side chains. GRASP integrates comparative protein composition data with annotation data from multiple public databases. Currently, GRASP includes information on proteins of 12 sequenced Drosophila (fruit fly) proteomes, which will be expanded to include increasingly diverse organisms over time. In this paper we illustrate the potential of GRASP for testing stoichioproteomic hypotheses by conducting an exploratory investigation into the composition of 12 Drosophila proteomes, testing the prediction that protein atomic content is associated with species ecology and with protein expression levels.
Results
Elements varied predictably along multivariate axes. Species were broadly similar, with the D. willistoni proteome a clear outlier. As expected, individual protein atomic content within proteomes was influenced by protein function and amino acid biochemistry. Evolution in elemental composition across the phylogeny followed less predictable patterns, but was associated with broad ecological variation in diet. Using expression data available for D. melanogaster, we found evidence consistent with selection for efficient usage of elements within the proteome: as expected, nitrogen content was reduced in highly expressed proteins in most tissues, most strongly in the gut, where nutrients are assimilated, and least strongly in the germline.
Conclusions
The patterns identified here using GRASP provide a foundation on which to base future research into the evolution of atomic composition in Drosophila and other taxa.
Nutrient recycling by fish can be an important part of nutrient cycles in both freshwater and marine ecosystems. As a result, understanding the mechanisms that influence excretion elemental ratios of fish is of great importance to a complete understanding of aquatic nutrient cycles. As fish consume a wide range of diets that differ in elemental composition, stoichiometric theory can inform predictions about dietary effects on excretion ratios.
We conducted a meta-analysis to test the effects of diet elemental composition on consumption and nutrient excretion by fish. We examined the relationship between consumption rate and diet N : P across all laboratory studies and calculated effect sizes for each excretion metric to test for significant effects.
Consumption rate of N, but not P, was significantly negatively affected by diet N : P. Effect sizes of diet elemental composition on consumption-specific excretion N, P and N : P in laboratory studies were all significantly different from 0, but effect size for raw excretion N : P was not significantly different from zero in laboratory or field surveys.
Our results highlight the importance of having a mechanistic understanding of the drivers of consumer excretion rates and ratios. We suggest that more research is needed on how consumption and assimilation efficiency vary with N : P and in natural ecosystems in order to further understand mechanistic processes in consumer-driven nutrient recycling.
Here, focusing on calcium, phosphorus and nitrogen, we used nutritional geometry to analyse individual-based data on foraging and extraction efficiencies, and combined these with data on reproduction and migratory behaviour to understand how a large herbivorous carnivore can complete its life cycle on a narrow and seemingly low quality bamboo diet.
Behavioural results showed that pandas during the year switched between four main food categories involving the leaves and shoots of two bamboo species available. Nutritional analysis suggests that these diet shifts are related to the concentrations and balances of calcium, phosphorus and nitrogen. Notably, successive shifts in range use and food type corresponded with a transition to higher concentrations and/or a more balanced intake of these multiple key constituents.
Our study suggests that pandas obligatorily synchronize their seasonal migration and reproduction with the disjunct nutritional phenologies of two bamboo species. This finding has potentially important implications for habitat conservation for this species and, more generally, draws attention to the need for understanding the nutritional basis of food selection in devising management plans for endangered species.
Serial femtosecond crystallography requires reliable and efficient delivery of fresh crystals across the beam of an X-ray free-electron laser over the course of an experiment. We introduce a double-flow focusing nozzle to meet this challenge, with significantly reduced sample consumption, while improving jet stability over previous generations of nozzles. We demonstrate its use to determine the first room-temperature structure of RNA polymerase II at high resolution, revealing new structural details. Moreover, the double flow-focusing nozzles were successfully tested with three other protein samples and the first room temperature structure of an extradiol ring-cleaving dioxygenase was solved by utilizing the improved operation and characteristics of these devices.