Suchergebnisse

Protein turnover reflects the continual synthesis and breakdown of body proteins, and can be measured at a whole-body (i.e. aggregated across all body proteins) or tissue (e.g. skeletal muscle only) level using stable isotope methods. Evaluating protein turnover in free-living environments, such as military training, can help inform protein requirements. We undertook a narrative review of published literature with the aim of reviewing the suitability of, and advancements in, stable isotope methods for measuring protein turnover in field research. The 2 primary approaches for measuring protein turnover are based on precursor- and end-product methods. The precursor method is the gold-standard for measuring acute (over several hours) skeletal muscle protein turnover, whereas the end-product method measures chronic (over several weeks) skeletal muscle protein turnover and provides the opportunity to monitor free-living activities. Both methods require invasive procedures such as the infusion of amino acid tracers and muscle biopsies to assess the uptake of the tracer into tissue. However, the end-product method can also be used to measure acute (over 9–24 h) whole-body protein turnover noninvasively by ingesting (15)N-glycine, or equivalent isotope tracers, and collecting urine samples. The end-product method using (15)N-glycine is a practical method for measuring whole-body protein turnover in the field over short (24 h) time frames and has been used effectively in recent military field research. Application of this method may improve our understanding of protein kinetics during conditions of high physiological stress in free-living environments such as military training.

Synaptic proteins play an important role for the regulation of synaptic plasticity. Numerous studies have identified and revealed individual synaptic protein functions using protein overexpression or deletion. In neuropathic pain nociceptive stimuli conveyed from the periphery repetitively stimulate neurons in the central nerve system, brain and spinal cord. Neuronal activities change the turnover (synthesis and degradation) rate of synaptic proteins. Thus, the analysis of synaptic protein turnover rather than just expression level change is critical for studying the role of synaptic proteins in synaptic plasticity. Here, we analyzed synaptosomal proteome in the anterior cingulate cortex (ACC) to identify protein turnover rate changes caused by peripheral nerve injury. Whereas PKCγ levels were not altered, we found that the protein's turnover rate decreased after peripheral nerve injury. Our results suggest that postsynaptic PKCγ synthesized by neuronal activities in the ACC is translocated to the postsynaptic membrane with an extended half-life. ; This work was supported by two National Research Foundation (NRF) of Korea grants funded by the Korean government (MSIP) [NRF2012R1A3A1050385 to BKK and 2018R1C1B6008530 to HGK] and the Max Planck Society to CWT and DIP.

Differences in producing performance and organoleptic meat characteristics among pig genotypes and/or producing types are widely known. These parameters are also subjected to the animal's development, feeding and management. Detailed knowledge of the effects of production phase (PP), pig producing type (PT), dietary protein availability and their interactions on nutrient digestibility, nitrogen balance and protein metabolism is essential information to improve precision feeding techniques. The experiment was a 2 (PP) × 2 (PT) × 2 (diet) factorial design conducted with 32 male pigs, 16 entire F2 pigs progeny of Pietrain sires and Duroc × Landrace dams, and 16 castrated purebred Durocs belonging to two production phases (growing: 29.5 ± 3.19 v. fattening: 88.6 ± 6.26 kg BW), and assigned to one of two dietary CP levels, either standard (SP: 17% in growing and 15% in fattening) or low (LP: 15% in growing and 13% in fattening). Viscera and muscle fractional protein synthesis rates (FSRs; %/day) were conducted through a single infusion of 15% L-[ring-2H5]-phenylalanine, with subsequent blood sampling from 12 to 40 min, and sample collection of liver, duodenum, biceps femoris and longissimus dorsi skeletal muscles after sacrifice. Fattening animals acquired a greater feed ingestion capacity, average daily gain (P < 0.01) and apparent ileal digestibility, whereas growing pigs showed higher FSRs in both viscera (duodenum and liver) and in longissimus dorsi. F2 pigs showed higher average daily gain, nitrogen retention rates and FSR in liver and longissimus dorsi (P < 0.01). Nevertheless, apparent ileal digestibility in all essential amino acids was lower in F2 compared with Duroc pigs (P < 0.05). Protein metabolism was barely influenced by dietary CP content, although animals fed LP registered the lowest apparent ileal digestibility for CP and also for most of the essential amino acids compared with SP-fed pigs. This information may reveal differences in amino acid requirements between both PTs, with Duroc pigs receiving excess of dietary amino acids ; This study was a part of the Feed-a-Gene project and receivedfunding from the European Union's H2020 program under NationalInstitutes of Health (grant number 633531), as well as SpanishNational funding by MINECO (AGL2017-89289-R). L. Sarri is recip-ient of a research training grant from the Generalitat de Catalunya-European Social Funds (2019 FI_B 00416).

The focus of this book is on the role of protein metabolism and homeostasis in aging. An overview is provided of the current knowledge in the area, including protein synthesis, accuracy and repair, post-translational modifications, degradation and turnover, and how they define and influence aging. The chapters mainly focus on well-characterized factors and pathways, but new areas are also presented, where associations with aging are just being elucidated by current experimental data.

Precise control of protein turnover is essential for cellular homeostasis. The ubiquitin-proteasome system is well established as a major regulator of protein degradation, but an understanding of how inherent structural features influence the lifetimes of proteins is lacking. We report that yeast, mouse, and human proteins with terminal or internal intrinsically disordered segments have significantly shorter half-lives than proteins without these features. The lengths of the disordered segments that affect protein half-life are compatible with the structure of the proteasome. Divergence in terminal and internal disordered segments in yeast proteins originating from gene duplication leads to significantly altered half-life. Many paralogs that are affected by such changes participate in signaling, where altered protein half-life will directly impact cellular processes and function. Thus, natural variation in the length and position of disordered segments may affect protein half-life and could serve as an underappreciated source of genetic variation with important phenotypic consequences. ; HFSP RGY0073/2010 ; Dutch government FES0908 ; Netherlands Genomics Initiative 050-060-452 ; Herchel Smith Research Studentship Fund ; LMB International PhD Programme ; EMBO STF 312-2011 ; Hungarian Academy of Sciences LP2012-41/2012 ; FEBS FP7-PEOPLE-2012-IEF 299105 ; PrioNet Canada ; UBC ; NIH R01GM63004, U54GM105816 ; Welch Foundation F-1817 ; MRC ; EMBO Young Investigator Programme ; Trinity College ; ERASysBio+ ; Molecular Biosciences

Age-related macular degeneration (AMD) is one of the commonest causes of sight loss in the elderly population and to date there is no intervention that slows or prevents early AMD disease progressing to blinding neovascularization or geographic atrophy. AMD is a complex disease and factors proposed to contribute to the development and progression of disease include aging, genetics, epigenetics, oxidative stress, pro-inflammatory state, and life-style factors such as smoking, alcohol, and high fat diet. Here, we generate a knowledge repository of pathways and protein-protein interaction (PPI) networks likely to be implicated in AMD pathogenesis, such as complement activation, lipid trafficking and metabolism, vitamin A cycle, oxidative stress, proteostasis, bioenergetics, autophagy/mitophagy, extracellular matrix (ECM) turnover, and choroidal vascular dropout. Two disctinct clusters ermerged from the networks for parainflamation and ECM homeostasis, which may represent two different disease modules underlying AMD pathology. Our analyses also suggest that the disease manifests primarily in RPE/choroid and less in neural retina. The use of standardized syntax when generating maps of these biological processes (SBGN standard) and networks (PSI standard) enables visualization of complex information in graphical programs such as CellDesigner and Cytoscape and enhances reusability and extension of data. The ability to focus onto subnetworks, multiple visualizations and simulation options will enable the AMD research community to computationally model subnetworks or to test experimentally new hypotheses arising from connectivities in the AMD pathway map. ; This work has received funding from the European Union's Horizon 2020 research and innovation program under grant agreement No. 634479 (EYE-RISK—to P.J.L., L.S., and C.K.). This work was supported by the Spanish Ministry of Economy and Competitiveness, Plan Nacional BIO2012-39754 and the European Fund for Regional Development (to L.S.). L.S. acknowledges the support ...

Abstract
Precise arrangement of actin, myosin, and other regulatory components in a sarcomeric pattern is critical for producing contractile forces in striated muscles. Actin-interacting protein 1 (AIP1), also known as WD-repeat protein 1 (WDR1), is one of essential factors that regulate sarcomeric assembly of actin filaments. In the nematode Caenorhabditis elegans, mutation in unc-78, encoding one of the two AIP1 isoforms, causes severe disorganization of sarcomeric actin filaments and near paralysis, but mutation in sup-13 suppresses the unc-78-mutant phenotypes to restore nearly normal sarcomeric actin organization and worm motility. Here, we identified that sup-13 is a nonsense allele of arrd-15 encoding an α-arrestin. The sup-13/arrd-15 mutation suppressed the phenotypes of unc-78 null mutant but required aipl-1 that encodes a second AIP1 isoform. aipl-1 was normally expressed highly in embryos and downregulated in mature muscle. However, in the sup-13/arrd-15 mutant, the AIPL-1 protein was maintained at high levels in adult muscle to compensate for the absence of the UNC-78 protein. The sup-13/arrd-15 mutation caused accumulation of ubiquitinated AIPL-1 protein, suggesting that a normal function of sup-13/arrd-15 is to enhance degradation of ubiquitinated AIPL-1, thereby promoting transition of AIP1 isoforms from AIPL-1 to UNC-78 in developing muscle. These results suggest that α-arrestin is a novel factor to promote isoform turnover by enhancing protein degradation.

We evaluated the impact of protein supplementation on adaptations to arduous concurrent training in healthy adults with potential applications to individuals undergoing military training. Peer-reviewed papers published in English meeting the population, intervention, comparison and outcome criteria were included. Database searches were completed in PubMed, Web of science and SPORTDiscus. Study quality was evaluated using the COnsensus based standards for the selection of health status measurement instruments checklist. Of 11 studies included, nine focused on performance, six on body composition and four on muscle recovery. Cohen's d effect sizes showed that protein supplementation improved performance outcomes in response to concurrent training (ES = 0.89, 95% CI = 0.08–1.70). When analysed separately, improvements in muscle strength (SMD = +4.92 kg, 95% CI = −2.70–12.54 kg) were found, but not in aerobic endurance. Gains in fat-free mass (SMD = +0.75 kg, 95% CI = 0.44–1.06 kg) and reductions in fat-mass (SMD = −0.99, 95% CI = −1.43–0.23 kg) were greater with protein supplementation. Most studies did not report protein turnover, nitrogen balance and/or total daily protein intake. Therefore, further research is warranted. However, our findings infer that protein supplementation may support lean-mass accretion and strength gains during arduous concurrent training in physical active populations, including military recruits.

We evaluated the impact of protein supplementation on adaptations to arduous concurrent training in healthy adults with potential applications to individuals undergoing military training. Peer-reviewed papers published in English meeting the population, intervention, comparison and outcome criteria were included. Database searches were completed in PubMed, Web of science and SPORTDiscus. Study quality was evaluated using the COnsensus based standards for the selection of health status measurement instruments checklist. Of 11 studies included, nine focused on performance, six on body composition and four on muscle recovery. Cohen's d effect sizes showed that protein supplementation improved performance outcomes in response to concurrent training (ES = 0.89, 95% CI = 0.08–1.70). When analysed separately, improvements in muscle strength (SMD = +4.92 kg, 95% CI = −2.70–12.54 kg) were found, but not in aerobic endurance. Gains in fat-free mass (SMD = +0.75 kg, 95% CI = 0.44–1.06 kg) and reductions in fat-mass (SMD = −0.99, 95% CI = −1.43–0.23 kg) were greater with protein supplementation. Most studies did not report protein turnover, nitrogen balance and/or total daily protein intake. Therefore, further research is warranted. However, our findings infer that protein supplementation may support lean-mass accretion and strength gains during arduous concurrent training in physical active populations, including military recruits.

Green Fluorescent protein (GFP) from Aequorea ictoria was used as an in vivo reporter protein when fused to the C-terminus of the Jen1 lactate permease of Saccharomyces cerevisiae. The Jen1 protein tagged with GFP is a functional lactate transporter with a cellular abundance of 1670 molecules/cell, and a catalyticcentre activity of 123 s-1. It is expressed and tagged to the plasma membrane under induction conditions. The factors involved in proper localization and turnover of Jen1p were revealed by expression of the Jen1p-GFP fusion protein in a set of strains bearing mutations in specific steps of the secretory and endocytic pathways. The chimaeric protein Jen1p-GFP is targeted to the plasma membrane via a Sec6-dependent process; upon treatment with glucose, it is endocytosed via END3 and targeted for degradation in the vacuole. Experiments performed in a Ddoa4 mutant strain showed that ubiquitination is associated with the turnover of the permease. ; FEBS - short-term fellowship. ; Portugal - Programa Operacional "Ciência, Tecnologia, Inovação" (POCTI) - POCTI/1999/BME/36 625. ; Portuguese Government - PRAXIS ...

Not Available ; Introduction Photosystem II (PSII) protein complex plays an essential role in the entire photosynthesis process. Various known and unknown protein factors are involved in the dynamics of the PSII complex that need to be characterized in crop plants for enhancing photosynthesis efficiency and productivity. Objectives The experiments were conducted to decipher the regulatory proteins involved in PSII dynamics of rice crop. Methods A novel rice regulatory protein PAP90 (PSII auxiliary protein ~90 kDa) was characterized by generating a loss-of-function mutant pap90. The mutation was characterized at molecular level followed by various experiments to analyze the morphological, physiological and biochemical processes of mutant under control and abiotic stresses. Results The pap90 mutant showed reduced photosynthesis due to D1 protein instability that subsequently causes inadequate accumulation of thylakoid membrane complexes, especially PSII and decreases PSII functional efficiency. Expression of OsFtsH family genes and proteins were induced in the mutant, which are known to play a key role in D1 protein degradation and turnover. The reduced D1 protein accumulation in the mutant increased the production of reactive oxygen species (ROS). The accumulation of ROS along with the increased activity of antioxidant enzymes and induced expression of stress-associated genes and proteins in pap90 mutant contributed to its water-limited stress tolerance ability. Conclusion We propose that PAP90 is a key auxiliary protein that interacts with D1 protein and maintains its stability, thereby promoting subsequent assembly of the PSII and associated membrane complexes. ; Department of Biotechnology, Government of India FileNo. BT/PR-13105/AGR/02/684/2009.