Phytoplankton stoichiometry

Because phytoplankton live at the interface between the abiotic and the biotic compartments of ecosystems, they play an important role in coupling multiple nutrient cycles. The quantitative details of how these multiple nutrient cycles intersect is determined by phytoplankton stoichiometry. Here we review some classic work and recent advances on the determinants of phytoplankton stoichiometry and their role in determining ecosystem stoichiometry. First, we use a model of growth with flexible stoichiometry to reexamine Rhee and Goldman’s classic chemostat data. We also discuss a recent data compilation by Hall and colleagues that illustrates some limits to phytoplankton flexibility, and a model of physiological adaptation that can account for these results. Second, we use a model of resource allocation to determine the how the optimal nitrogen-to-phosphorus stoichiometry depends on the ecological conditions under which species grow and compete. Third, we discuss Redfield’s mechanism for the homeostasis of the oceans’ nitrogen-to-phosphorus stoichiometry and show its robustness to additional factors such as iron-limitation and temporal fluctuations. Finally, we suggest areas for future research.     

Nutrient stoichiometry shapes microbial coevolution

Coevolution is a force contributing to the generation and maintenance of biodiversity. It is influenced by environmental conditions including the scarcity of essential resources, which can drive the evolution of defence and virulence traits. We conducted a long‐term chemostat experiment where the marine cyanobacterium Synechococcus was challenged with a lytic phage under nitrogen (N) or phosphorus (P) limitation. This manipulation of nutrient stoichiometry altered the stability of host–parasite interactions and the underlying mode of coevolution. By assessing the infectivity with > 18 000 pairwise challenges, we documented directional selection for increased phage resistance, consistent with arms‐race dynamics while phage infectivity fluctuated through time, as expected when coevolution is driven by negative frequency‐dependent selection. The resulting infection networks were 50% less modular under N‐ versus P‐limitation reflecting host‐range contraction and asymmetric coevolutionary trajectories. Nutrient stoichiometry affects eco‐evolutionary feedbacks in ways that may alter the dynamics and functioning of environmental and host‐associated microbial communities.     

Ordinary stoichiometry of extraordinary microorganisms

All life on Earth seems to be made of the same chemical elements in relatively conserved proportions (stoichiometry). Whether this stoichiometry is conserved in settings that differ radically in physicochemical conditions (extreme environments) from those commonly encountered elsewhere on the planet provides insight into possible stoichiometries for putative life beyond Earth. Here, we report measurements of elemental stoichiometry for extremophile microbes from hot springs of Yellowstone National Park (YNP). Phototrophic and chemotrophic microbes were collected in locations spanning large ranges of temperature (24 °C to boiling), pH (1.6–9.6), redox (0.1–7.2 mg L^?1 dissolved oxygen), and nutrient concentrations (0.01–0.25 mg L^?1 , 0.7–12.9 mg L^?1 , 0.01–42 mg L^?1 NH₄⁺, 0.003–1.1 mg L^?1 P mostly as phosphate). Despite these extreme conditions, the microbial cells sampled had a major and trace element stoichiometry within the ranges commonly encountered for microbes living in the more moderate environments of lakes and surface oceans. The cells did have somewhat high C:P and N:P ratios that are consistent with phosphorus (P) limitation. Furthermore, chemotrophs and phototrophs had similar compositions with the exception of Mo content, which was enriched in cells derived from chemotrophic sites. Thus, despite the extraordinary physicochemical and biological diversity of YNP environments, life in these settings, in a stoichiometric sense, remains much the same as we know it elsewhere.     

Alcohol-induced changes of beta-lactoglobulin-retinol-binding stoichiometry

It has been demonstrated using CD that ethanol induces important secondary structure changes of beta-lactoglobulin. CD spectra indicate that beta-lactoglobulin secondary structure, which is mainly composed of beta-strands, becomes mostly alpha-helical under the influence of the solvent polarity changes. The midpoint of beta-strand/alpha-helix transition in beta-lactoglobulin is observed at dielectric constant approximately 60 (35% ethanol; v/v). According to CD measurements, the ethanol-dependent secondary structure changes are reversible. The alkylation of lysines epsilon-NH2 in beta-lactoglobulin weakens the central beta-barrel structure, since the beta-strand/alpha-helix transition midpoint of alkylated beta-lactoglobulin is shifted to lower ethanol concentration (25% ethanol; v/v). beta-Lactoglobulin structural changes are triggering the dissociation of the beta-lactoglobulin-retinol complex as judged from complete quenching of its fluorescence in ethanol concentration greater than 30% (v/v). However, in 20% ethanol (v/v), beta-lactoglobulin still retains most of its native secondary structure as shown by CD and, in this condition, one beta-lactoglobulin molecule binds an additional second retinol molecule. This suggests that the highly populated species observed around 20% ethanol (v/v) might represent an intermediate state able to bind two molecules of retinol.     

Consumer-resource stoichiometry in detritus-based streams

Stoichiometric relationships between consumers and resources in detritus‐based ecosystems have received little attention, despite the importance of detritus in most food webs. We analysed carbon (C), nitrogen (N), and phosphorus (P) content of invertebrate consumers, and basal food resources in two forested headwater streams (one reference and the other nutrient‐enriched). We found large elemental imbalances between consumers and food resources compared with living plant‐based systems, particularly in regard to P content, which were reduced with enrichment. Enrichment significantly increased nutrient content of food resources (consistent with uptake of N and P by detritus‐associated microbes). P content of some invertebrates also increased in the enriched vs. reference stream, suggesting deviation from strict homeostasis. Nutrient content varied significantly among invertebrate functional feeding groups, orders and, to some extent, size classes. Future application of stoichiometric theory to detritus‐based systems should consider the potential for relatively large consumer‐resource elemental imbalances and P storage by insect consumers.     

Epithelial Na+ channel subunit stoichiometry

Ion channels, including the epithelial Na(+) channel (ENaC), are intrinsic membrane proteins comprised of component subunits. Proper subunit assembly and stoichiometry are essential for normal physiological function of the channel protein. ENaC comprises three subunits, alpha, beta, and gamma, that have common tertiary structures and much amino acid sequence identity. For maximal ENaC activity, each subunit is required. The subunit stoichiometry of functional ENaC within the membrane remains uncertain. We combined a biophysical approach, fluorescence intensity ratio analysis, used to assess relative subunit stoichiometry with total internal reflection fluorescence microscopy, which enables isolation of plasma membrane fluorescence signals, to determine the limiting subunit stoichiometry of ENaC within the plasma membrane. Our results demonstrate that membrane ENaC contains equal numbers of each type of subunit and that at steady state, subunit stoichiometry is fixed. Moreover, we find that when all three ENaC subunits are coexpressed, heteromeric channel formation is favored over homomeric channels. Electrophysiological results testing effects of ENaC subunit dose on channel activity were consistent with total internal reflection fluorescence/fluorescence intensity ratio findings and confirmed preferential formation of heteromeric channels containing equal numbers of each subunit.     

Mechanistic stoichiometry of mitochondrial oxidative phosphorylation

P/O ratios of rat liver mitochondria were measured with particular attention to systematic errors. Corrections for energy loss during oxidative phosphorylation were made by measurement of respiration as a function of mitochondrial membrane potential. The corrected values were close to 1, 0.5, and 1 at the three coupling sites, respectively. These values are consistent with recent measurements of mitochondrial proton transport.     

The temperature-dependent stoichiometry of mixed cryoimmunoglobulins

The interaction of three monoclonal rheumatoid factor IgM molecules with IgG antigens has been studied utilizing immunoglobulins isolated from three mixed cryoglobulins. Static light scattering measurements show that the stoichiometry of these immune complexes changes in a temperature-dependent manner from IgM(IgG)0-2 at temperatures greater than 37 degrees C to IgM(IgG)5 complexes at temperatures below 15 degrees C. These results were confirmed by the analysis of the composition of polyethyleneglycol-precipitated complexes. For one mixed cryoglobulin (Glo), temperature-dependent changes in stoichiometry were also verified by chemical cross-linking studies. Binding constants were determined by Scatchard analysis of light scattering data and by fluorescence polarization measurements. Values on the order of 10(5) M-1 were obtained for three monoclonal rheumatoid factor IgM molecules. Glo was further investigated by dynamic light scattering and partial specific volume measurements. Both dynamic light scattering and partial specific volume measurements provided evidence for surprising shape changes of the IgM X IgG complex as a function of temperature and IgG stoichiometry. Collectively, the data support the simple hypothesis that cryoprecipitation of mixed cryoglobulins occurs as a consequence of increases in the size (stoichiometry) of the complexes that are formed at low temperatures.     

植物生态化学计量内稳性特征

化学计量内稳性是生态化学计量学研究的核心概念之一,是指生物在面对外界变化的时候保持自身化学组成相对稳定的能力,其反映了生物对周围环境变化作出的生理和生化响应与适应。通过研究植物生态化学计量内稳性,有助于深入了解植物对环境的适应策略和生态适应性,以及植物化学计量内稳性与生态系统功能的关系,但目前关于植物生态化学计量内稳性的研究较少。已有的研究结果表明:不同物种或功能群由于其生长策略不同而具有不同的生态化学计量内稳性特征;同一物种的不同器官、不同生长阶段以及不同元素的内稳性存在较大的差异。该文对植物生态化学计量内稳性概念、内稳性指数的测算方法,不同植物物种或功能群、不同器官、不同生长阶段内稳性特征,以及植物内稳性与生态系统结构、功能和稳定性的关系等方面进行了综述,并结合现已开展的工作,对有待进一步拓展的相关植物生态化学计量内稳性研究领域进行了展望,以期为促进国内相关研究工作的开展提供参考。     

FRET-based analysis of TRPC subunit stoichiometry

By analogy to other cation channel subunits with six transmembrane-spanning domains, the seven members of the "classical" or "canonical" transient receptor potential channels (TRPC) family are believed to assemble into homo- or heterotetrameric complexes. These complexes have been verified by classical methods such as coimmunoprecipitation, crosslinking analysis or functional assays applying dominant negative pore mutants. More recently, fluorescence resonance energy transfer (FRET)-a measure for the close proximity of fluorescent molecules-has become instrumental in monitoring protein assembly in living cells. Here we demonstrate further possibilities and verification procedures of the FRET technology to test the assembly of ion channel subunits. Temporally and spatially resolved FRET imaging demonstrates an early assembly of TRPC subunits in the endoplasmic reticulum and the Golgi apparatus. Confocal FRET imaging verifies FRET signals over the plasma membrane at high spatial resolution. Taking advantage of the quantitative analysis of digital video imaging, we demonstrate that FRET between TRPC subunits is only poorly concentration-dependent. Moreover, a correlation between the efficiency of energy transfer and the molar ratio of the FRET donor to the acceptor was exploited to verify the tetrameric stoichiometry of TRPC complexes. Finally, we introduce a competition-FRET assay to test the ability of wild-type TRPC subunits to recruit fluorescent TRPC subunits into separate channel complexes.     

O-GlcNAc glycosylation stoichiometry of the FET protein family: only EWS is glycosylated with a high stoichiometry

Of the FET (fused in sarcoma [FUS]/Ewing sarcoma protein [EWS]/TATA binding protein-associated factor 15 [TAF15]) family of heterogeneous nuclear ribonucleoprotein particle proteins, FUS and TAF15 are consistently and EWS variably found in inclusion bodies in neurodegenerative diseases such as frontotemporal lobar degeneration associated with FUS. It is speculated that dysregulation of FET proteins at the post-translational level is involved in their cytoplasmic deposition. Here, the O-linked β-N-acetylglucosamine (O-GlcNAc) glycosylation stoichiometry of the FET proteins was chemoenzymatically analyzed, and it was found that only EWS is dynamically glycosylated with a high stoichiometry in the neural cell lines tested and in mouse brain. It was also confirmed that EWS, but not FUS and TAF15, is glycosylated with a high stoichiometry not only in the neural cells but also in the non-neural cell lines tested. These results indicate that O-GlcNAc glycosylation imparts a physicochemical property on EWS that is distinct from that of the other FET proteins in most of cell lineages or tissues.     

Optimal metabolic regulation along resource stoichiometry gradients

Most heterotrophic organisms feed on substrates that are poor in nutrients compared to their demand, leading to elemental imbalances that may constrain their growth and function. Flexible carbon (C)‐use efficiency (CUE, C used for growth over C taken up) can represent a strategy to reduce elemental imbalances. Here, we argue that metabolic regulation has evolved to maximise the organism growth rate along gradients of nutrient availability and translated this assumption into an optimality model that links CUE to substrate and organism stoichiometry. The optimal CUE is predicted to decrease with increasing substrate C‐to‐nutrient ratio, and increase with nutrient amendment. These predictions are generally confirmed by empirical evidence from a new database of c. 2200 CUE estimates, lending support to the hypothesis that CUE is optimised across levels of organisation (microorganisms and animals), in aquatic and terrestrial systems, and when considering nitrogen or phosphorus as limiting nutrients.