Function and dynamics of the intrinsically disordered carboxyl terminus of ß2 adrenergic receptor.

Advances in structural biology have provided important mechanistic insights into signaling by the transmembrane core of G-protein coupled receptors (GPCRs); however, much less is known about intrinsically disordered regions such as the carboxyl terminus (CT), which is highly flexible and not visible in GPCR structures. The ß2 adrenergic receptor's (ß2AR) 71 amino acid CT is a substrate for GPCR kinases and binds ß-arrestins to regulate signaling. Here we show that the ß2AR CT directly inhibits basal and agonist-stimulated signaling in cell lines lacking ß-arrestins. Combining single-molecule fluorescence resonance energy transfer (FRET), NMR spectroscopy, and molecular dynamics simulations, we reveal that the negatively charged ß2AR-CT serves as an autoinhibitory factor via interacting with the positively charged cytoplasmic surface of the receptor to limit access to G-proteins. The stability of this interaction is influenced by agonists and allosteric modulators, emphasizing that the CT plays important role in allosterically regulating GPCR activation.

[Change in nitrogen availability of northern peatlands and its effect on carbon sink function]. / åŒ—æ–¹æ³¥ç‚­åœ°æ°®ç´ æœ‰æ•ˆæ€§çš„å˜åŒ–åŠå ¶å¯¹ç¢³æ±‡åŠŸèƒ½çš„å½±å“.

Northern peatlands are typical nitrogen-limited ecosystems, which are sensitive to global climate change and human activities. The increases of endogenous available nitrogen caused by climate warming and exogenous nitrogen input caused by human activities changed the nitrogen availability of northern peatlands, and would affect carbon and nitrogen cycling and carbon sink function of peatland. Here, we review the influence factors of carbon accumulation rate and carbon sink function in northern peatlands. The effects of nitrogen deposition, freezing and thawing, fire and other factors on nitrogen availability of northern peatlands were reviewed. The responses of plants and soil microorganisms to changes in nitrogen availability were elaborated from carbon fixation and carbon emission processes, respectively. The research related to carbon sink function of peat ecosystems under the influence of glo-bal change was prospected, aiming to help the implementation of the 'double carbon' goal.

Warming-Induced Labile Carbon Change Soil Organic Carbon Mineralization and Microbial Abundance in a Northern Peatland.

Climate warming affects the carbon cycle of northern peatlands through temperature rises and a changing carbon availability. To clarify the effects of elevated temperature and labile carbon addition on SOC mineralization, as well as their microbial driving mechanisms, topsoil (0-10 cm) and subsoil (10-20 cm) were collected from a peatland in the Great Hing'an Mountains and incubated with or without 13C-glucose at 10 °C and 15 °C for 42 days. The results showed that 5 °C warming significantly stimulated SOC mineralization along with NH4+-N and NO3--N content increases, as well as a decrease in invertase and urease activities. Glucose addition triggered a positive priming effect (PE) in the early stage of the incubation but changed to a negative PE in the late stage of the incubation. Glucose likely regulates carbon dynamics by altering fungi: bacteria, soil invertase, and ß-glucodase activities, and MBC, DOC, NH4+-N contents. Glucose addition increased fungal abundance in 0-10 cm at 10 °C and 15 °C, and 10-20 cm at 10 °C, respectively, but significantly decreased fungal abundance in 10-20 cm at 15 °C. Glucose addition decreased bacterial abundance in 0-10 cm at 10 °C but increased bacterial abundance in 10-20 cm soil at 10 °C, and in 0-10 and 10-20 cm soils at 15 °C, respectively. Glucose addition significantly decreased the fungi: bacteria ratio in 0-20 cm soils at 15 °C. In addition, Q10 was significantly positively correlated with the changes in soil DOC, NH4+-N contents, invertase, and ß-glucosidase activities, while negatively correlated with fungi: bacteria and urease activities after 5 °C of warming, and glucose addition significantly increased the Q10. Labile carbon may decrease carbon losses in northern peatlands that inhibit warming-induced carbon emission increase, thus partially buffering soil carbon content against change.

Long-term nitrogen addition alters peatland plant community structure and nutrient resorption efficiency.

As an elemental carbon (C) and nitrogen (N) pool in the world, peatlands are very sensitive to environmental changes. Under global warming, the increase in available N affects the dynamic changes of plant community structure and nutrients in a permafrost peatland. This study was based on a long-term in situ N addition experiment that had been conducted for 9 years. It utilized the peatland in the permafrost area of Great Hing'an Mountain as the research object to analyze the effects of N addition on the growth characteristics, community structure, and nutrient dynamics of peatland plants. The N inputs were N1: 6 g N m-2·year-1, N2: 12 g N m-2·year-1 and N3: 24 g N m-2·year-1, respectively. Our results showed that the adding N can affect the plant community structure of peatland by affecting the plant growth characteristics. The diversity and richness of plant species in the peatland decreased as the concentration of added N increased. The long-term N addition can reduce the N limitation of plants to some extent. Still, it could further aggravate their phosphorus (P) limitation, resulting in the joint limitation of N and P or the complete limitation by P. The N resorption efficiency decreased with the increase of N addition level. The P resorption efficiency of different plants had varied responses to the changes in the N nutrient environment. Our study clarified the impact of long-term N addition on the plant community structure and nutrient dynamics of peatland in a permafrost area and provided an important theoretical basis to accurately evaluate the carbon and nitrogen balance of peatland in a permafrost area owing to future climate change.

Effect of Nitrogen Addition on Soil Microbial Functional Gene Abundance and Community Diversity in Permafrost Peatland.

Nitrogen is the limiting nutrient for plant growth in peatland ecosystems. Nitrogen addition significantly affects the plant biomass, diversity and community structure in peatlands. However, the response of belowground microbe to nitrogen addition in peatland ecosystems remains largely unknown. In this study, we performed long-term nitrogen addition experiments in a permafrost peatland in the northwest slope of the Great Xing'an Mountains. The four nitrogen addition treatments applied in this study were 0 g N·m-2·year-1 (CK), 6 g N·m-2·year-1 (N1), 12 g N·m-2·year-1 (N2), and 24 g N·m-2·year-1 (N3). Effects of nitrogen addition over a period of nine growing seasons on the soil microbial abundance and community diversity in permafrost peatland were analyzed. The results showed that the abundances of soil bacteria, fungi, archaea, nitrogen-cycling genes (nifH and b-amoA), and mcrA increased in N1, N2, and N3 treatments compared to CK. This indicated that nitrogen addition promoted microbial decomposition of soil organic matter, nitrogen fixation, ammonia oxidation, nitrification, and methane production. Moreover, nitrogen addition altered the microbial community composition. At the phylum level, the relative abundance of Proteobacteria increased significantly in the N2 treatment. However, the relative abundances of Actinobacteria and Verrucifera in the N2 treatment and Patescibacteria in the N1 treatment decreased significantly. The heatmap showed that the dominant order composition of soil bacteria in N1, N2, and N3 treatments and the CK treatment were different, and the dominant order composition of soil fungi in CK and N3 treatments were different. The N1 treatment showed a significant increase in the Ace and Chao indices of bacteria and Simpson index of fungi. The outcomes of this study suggest that nitrogen addition altered the soil microbial abundance, community structure, and diversity, affecting the soil microbial carbon and nitrogen cycling in permafrost peatland. The results are helpful to understand the microbial mediation on ecological processes in response to N addition.

Analysis of ß2AR-Gs and ß2AR-Gi complex formation by NMR spectroscopy.

The ß2-adrenergic receptor (ß2AR) is a prototypical G protein-coupled receptor (GPCR) that preferentially couples to the stimulatory G protein Gs and stimulates cAMP formation. Functional studies have shown that the ß2AR also couples to inhibitory G protein Gi, activation of which inhibits cAMP formation [R. P. Xiao, Sci. STKE 2001, re15 (2001)]. A crystal structure of the ß2AR-Gs complex revealed the interaction interface of ß2AR-Gs and structural changes upon complex formation [S. G. Rasmussen et al., Nature 477, 549-555 (2011)], yet, the dynamic process of the ß2AR signaling through Gs and its preferential coupling to Gs over Gi is still not fully understood. Here, we utilize solution nuclear magnetic resonance (NMR) spectroscopy and supporting molecular dynamics (MD) simulations to monitor the conformational changes in the G protein coupling interface of the ß2AR in response to the full agonist BI-167107 and Gs and Gi1 These results show that BI-167107 stabilizes conformational changes in four transmembrane segments (TM4, TM5, TM6, and TM7) prior to coupling to a G protein, and that the agonist-bound receptor conformation is different from the G protein coupled state. While most of the conformational changes observed in the ß2AR are qualitatively the same for Gs and Gi1, we detected distinct differences between the ß2AR-Gs and the ß2AR-Gi1 complex in intracellular loop 2 (ICL2). Interactions with ICL2 are essential for activation of Gs These differences between the ß2AR-Gs and ß2AR-Gi1 complexes in ICL2 may be key determinants for G protein coupling selectivity.

L. plantarum, L. fermentum, and B. breve Beads Modified the Intestinal Microbiota and Alleviated the Inflammatory Response in High-Fat Diet-Fed Mice.

This paper aims to study the effects of compound microbe-based beads on changes in the intestinal microbiota and alleviation of high-fat (HF) diet-induced inflammatory responses. Forty-eight mice were fed base chow or a high-fat diet for 4 weeks and then randomly separated into six groups: normal diet (group A), high-fat diet (group B), high-fat positive control (fed with high-fat chow plus Tetrahydrolipstatin, group C), high-fat chow plus B. breve beads (group D), high-fat chow plus L. plantarum-L. fermentum beads (group E), and high-fat chow plus L. plantarum-L. fermentum-B. breve beads (group F). The body weights were measured. The serum cytokine and lipid levels were determined by ELISA, and high-throughput sequence analysis of the fecal microbiota was conducted. Beads with cell encapsulation rates higher than 99% decreased the body weight from 50.97 ± 3.44 g in group B to 42.64 ± 2.63 g in group F at the end of the experiment (p = 0.00019). The total cholesterol content in group F was 80.14 ± 9.37 mmol/L, which was significantly lower than that in group A (96.13 ± 24.07 mmol/L) (p = 0.02765), group B (102.52 ± 12.20 mmol/L) (p = 0.00196), and group C (98.99 ± 11.32 mmol/L) (p = 0.00804). In addition, the serum IL-6 level showed no significant difference between group F and the base chow control group. The microbial cell-loaded bead intervention led to increased abundances of Bifidobacterium and Lactobacillus in mouse feces. Oral administration of three strain-based beads led to alleviation of inflammatory reactions in high-fat diet-fed mice.

Antifungal activity of 1-methylcyclopropene (1-MCP) against anthracnose (Colletotrichum gloeosporioides) in postharvest mango fruit and its possible mechanisms of action.

Anthracnose caused by Colletotrichum gloeosporioides is one of the most important postharvest diseases in mango fruit, often causing huge economic losses. In this study, the effect of 1-methylcyclopropene (1-MCP) against anthracnose in postharvest mango fruit and the mechanisms involved were investigated. 1-MCP induced reactive oxygen species (ROS) generation, damaged the mitochondria and destroyed the integrity of plasma membrane of spores of C. gloeosporioides, significantly suppressing spore germination and mycelial growth of C. gloeosporioides. 1-MCP also decreased the decay incidence and lesion expansion of mango fruit caused by C. gloeosporioides. For the first time this study demonstrated that 1-MCP suppressed anthracnose of postharvest mango fruit by directly inhibiting spore germination and mycelial growth of C. gloeosporioides, thus providing a promising strategy for disease control.

Deep sequencing identifies tissue-specific microRNAs and their target genes involving in the biosynthesis of tanshinones in Salvia miltiorrhiza.

Salvia miltiorrhiza is one of the most popular traditional medicinal herbs in Asian nations. Its dried root contains a number of tanshinones, protocatechuic aldehyde, salvianolic acid B and rosmarinic, and is used for the treatment of various diseases. The finding of microRNAs (miRNAs) and their target genes will help understand their biological role on the biosynthesis of tanshinones in S. miltiorrhiza. In the present study, a total of 452 known miRNAs corresponding to 589 precursor miRNAs (pre-miRNAs), and 40 novel miRNAs corresponding to 24 pre-miRNAs were identified in different tissues of S. miltiorrhiza by high-throughput sequencing, respectively. Among them, 62 miRNAs express only in root, 95 miRNAs express only in stem, 19 miRNAs express only in leaf, and 71 miRNAs express only in flower, respectively. By the degradome analysis, 69 targets potentially cleaved by 25 miRNAs were identified. Among them, acetyl-CoA C-acetyltransferase was cleaved by miR5072, and involved in the biosynthesis of tanshinones. This study provided valuable information for understanding the tissue-specific expression patterns of miRNAs in S. miltiorrhiza, and offered a foundation for future studies of the miRNA-mediated biosynthesis of tanshinones.

Toward an improved discrimination of outer membrane proteins using a sequence-based approach.

This article offers a novel sequence-based approach to discriminate outer membrane proteins (OMPs). The first step is to use a new representation approach, factor analysis scales of generalized amino acid information (FASGAI) representing hydrophobicity, alpha and turn propensities, bulky properties, compositional characteristics, local flexibility and electronic properties, etc., to characterize sequences of OMPs and non-OMPs. The subsequent data is then transformed into a uniform matrix by the auto cross covariance (ACC). The second step is to develop discrimination predictors of OMPs from non-OMPs using a support vector machine (SVM). The SVM predictors thus successfully produce a high Matthews correlation coefficient (MCC) of 0.916 on 208 OMPs from non-OMPs including 206 α-helical membrane proteins and 673 globular proteins by a fivefold cross validation test. Meanwhile, overall MCC values of 0.923 and 0.930 are obtained for the discrimination OMPs from the α-helical membrane proteins and the globular proteins, respectively. The results demonstrate that the FASGAI-ACC-SVM combination approach shows great prospect of application in the field of bioinformatics or proteomics studies.