The Structure of Detoxin D1

The structure of detoxin D₁, one of the main active principles of detoxio complex, has been established on the basis of the degradative studies and spectral evidences as depicted in formula (I).

Detoxin D₁ has been demonstrated to belong to a new class of the depsipeptide contained an amino acid designated detoxinine which was newly isolated as a natural product.     

Improved metabolic action of a bacterial lysine decarboxylase gene in tobacco hairy root cultures by its fusion to arbcS transit peptide coding sequence

The gene of a bacterial lysine decarboxylase (ldc) fused to arbcS transit peptide coding sequence (tp), and under the control of the CaMV 35S promoter, was expressed in hairy root cultures ofNicotiana tabacum. The fusion of theldc to the targeting signal sequence improved the performance of the bacterial gene in the plant cells in many respects. Nearly all transgenic hairy root cultures harbouring the35S-tp-ldc gene contained distinctly higher lysine decarboxylase activity (from 1.5 to 30 pkat LDC per mg protein) than those which had been transformed with constructs in which the gene had been directly cloned behind the CaMV 35S promoter. The higher enzyme activity led to the accumulation of up to 0.7% cadaverine on a dry mass basis. In addition, part of the cadaverine pool was used for increased biosynthesis of anabasine, an alkaloid which was hardly detectable in control cultures. The best line contained anabasine levels of 0.5% dry mass, which could further be enhanced by feeding of lysine.     

Isolation and Characterization of Strains Defective in Vacuolar Ornithine Permease inNeurospora crassa

Vacuolar uptake of ornithine and lysine was characterized inNeurospora crassausing a cupric ion permeabilization system. Michaelis constants were measured as 1.4 mM for lysine and 11.0 mM for ornithine, and maximal velocities were determined. Vacuolar lysine uptake was shown to be inhibited competitively byl-arginine and histidine while ornithine uptake was inhibited by a variety of amino acids. Strains defective in the vacuolar ornithine permease were isolated using a filtration enrichment method. Two isolates—RSC-39 and RSC-63—had a reduced ability to accumulate ornithine. Vacuolar uptake of amino acids was measured using cupric ion-permeabilized mycelia; both strains had reduced ornithine uptake while lysine uptake and arginine uptake were normal. For both isolates, both the Michaelis constant and the maximal velocity for ornithine uptake were reduced compared to those of wild type. These results suggest that both strains are defective in the gene which encodes the vacuolar ornithine permease.     

Effects of castration age,dietary protein level and lysine/methionine ratio on animal performance,carcass and meat quality of Friesian steers intensively reared

The effects of castration age, dietary protein level and the dietary lysine/methionine (lys/met) ratio on animal performance, carcass characteristics and meat quality were studied in 64 intensively reared Friesian steers. Animals underwent castration procedures at 15 days old or at 5 months old. Dietary treatments started at 90 days old, with eight animals from each castration age randomly allocated to each treatment: 14.6% v. 16.8% CP (DM basis), and 3.0 v. 3.4 lys/met, on a 2×2×2 design. The recommended ratio of 3.0 was reached with supplementation of protected methionine. Steers were slaughtered at 443.5±26.2 kg live weight when they reached 12 months old approximately. Average daily gain, cold carcass weight or carcass classification were not affected by any studied effect. Muscle moisture (P=0.024), C18:2n-6 percentage (P=0.047), polyunsaturated fatty acid/saturated fatty acid (P=0.049) and n-6/n-3 (P=0.003) were higher in late castrated animals. Both high levels of dietary protein (P=0.008) and lys/met ratio (P=0.048) increased the percentage of muscle in the carcass. A level of 16.8% of CP in the diet also increased the percentage of monounsaturated fatty acids in the intramuscular fat (P=0.032), whereas a ratio lys/met of 3.4 decreased the percentage of saturated fatty acids (P=0.028). Thus, it is recommended using diets with a high protein level (16.8%) and a high lys/met ratio (3.4) in animals slaughtered at a young age, in order to obtain carcasses with high muscle content without negatively affecting productive traits or intramuscular fat composition.     

Improved methods for targeting epigenetic reader domains of acetylated and methylated lysine

Responsible for interpreting histone post-translational modifications, epigenetic reader proteins have emerged as novel therapeutic targets for a wide range of diseases. Chemical probes have been critical in enabling target validation studies and have led to translational advances in cancer and inflammation-related pathologies. Here, we present the most recently reported probes of reader proteins that recognize acylated and methylated lysine. We will discuss challenges associated with achieving potent antagonism of reader domains and review ongoing efforts to overcome these hurdles, focusing on targeting strategies including the use of peptidomimetic ligands, allosteric modulators, and protein degraders.     

A current view on Tau protein phosphorylation in Alzheimer's disease

The functions of the neuronal microtubule-associated protein Tau in the central nervous system are regulated by manifold posttranslational modifications at more than 50 sites. Tau in healthy neurons carries multiple phosphate groups, mostly in its microtubule assembly domain. Elevated phosphorylation and aggregation of Tau are widely considered pathological hallmarks in Alzheimer’s disease (AD) and other tauopathies, triggering the quest for Tau posttranslational modifications in the disease context. However, the phosphorylation patterns of physiological and pathological Tau are surprisingly similar and heterogenous, making it difficult to identify specific modifications as therapeutic targets and biomarkers for AD. We present a concise summary of - and view on - important previous and recent advances in Tau phosphorylation analysis in the context of AD.     

Substrate Locking Promotes Dimer-Dimer Docking of an Enzyme Antibiotic Target

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Effective cancer therapy based on selective drug delivery into cells across their membrane using receptor-mediated endocytosis

Cancer is one of the major causes of death globally. The current treatment options are insufficient, leading to unmet medical needs in cancer treatment. Off-target side effects, multidrug resistance, selective distribution to cancerous tissues, and cell membrane permeation of anti-cancer agents are critical problems to overcome. There is a method to solve these problems by using receptor-mediated endocytosis (RME). It is well known that proteins such as integrin, HER2, EGFR, or other cancer biomarkers are specifically overexpressed on the surface of target cancer cells. By taking advantage of such specific receptors, payloads can be transported into cells through endocytosis using a conjugate composed of the corresponding ligands connected to the payloads by an appropriate linker. After RME, the payloads released by endosomal escape into the cytoplasm can exhibit the cytotoxic activity against cancer cells. Cell-penetrating peptides (CPPs), tumor-homing peptides (THPs), and monoclonal antibodies (mAbs) are utilized as ligands in this system. Antibody drug conjugates (ADCs) based on RME have already been used to cure cancer. In addition to the canonical conjugate method, nanocarriers for spontaneous accumulation in cancer tissue due to enhanced permeability and retention (EPR) effect are extensively used. In this review, I introduce the possibilities and advantages of drug design and development based on RME for the treatment of cancer.     

Dose–response evaluation of the standardized ileal digestible tryptophan : lysine ratio to maximize growth performance of growing-finishing gilts under commercial conditions

Environmental regulations as well as economic incentives have resulted in greater use of synthetic amino acids in swine diets. Tryptophan is typically the second limiting amino acid in corn-soybean meal-based diets. However, using corn-based co-products emphasizes the need to evaluate the pig’s response to increasing Trp concentrations. Therefore, the objective of these studies was to evaluate the dose–response to increasing standardized ileal digestible (SID) Trp : Lys on growth performance of growing-finishing gilts housed under large-scale commercial conditions. Dietary treatments consisted of SID Trp : Lys of 14.5%, 16.5%, 18.0%, 19.5%, 21.0%, 22.5% and 24.5%. The study was conducted in four experiments of 21 days of duration each, and used corn-soybean meal-based diets with 30% distillers dried grains with solubles. A total of 1166, 1099, 1132 and 975 gilts (PIC 337×1050, initially 29.9±2.0 kg, 55.5±4.8 kg, 71.2±3.4 kg and 106.2±3.1 kg BW, mean±SD) were used. Within each experiment, pens of gilts were blocked by BW and assigned to one of the seven dietary treatments and six pens per treatment with 20 to 28 gilts/pen. First, generalized linear mixed models were fit to data from each experiment to characterize performance. Next, data were modeled across experiments and fit competing dose–response linear and non-linear models and estimate SID Trp : Lys break points or maximums for performance. Competing models included broken-line linear (BLL), broken-line quadratic and quadratic polynomial (QP). For average daily gain (ADG), increasing the SID Trp : Lys increased growth rate in a quadratic manner (P<0.02) in all experiments except for Exp 2, for which the increase was linear (P<0.001). Increasing SID Trp : Lys increased (P<0.05) feed efficiency (G : F) quadratically in Exp 1, 3 and 4. For, ADG the QP was the best fitting dose–response model and the estimated maximum mean ADG was obtained at a 23.5% (95% confidence interval (CI): [22.7, 24.3%]) SID Trp : Lys. For maximum G : F, the BLL dose–response models had the best fit and estimated the SID Trp : Lys minimum to maximize G : F at 16.9 (95% CI: [16.0, 17.8%]). Thus, the estimated SID Trp : Lys for 30 to 125 kg gilts ranged from a minimum of 16.9% for maximum G : F to 23.5% for maximum ADG.     

Two‐domain analysis of JmjN‐JmjC and PHD‐JmjC lysine demethylases: Detecting an inter‐domain evolutionary stress

Residues at different positions of a multiple sequence alignment sometimes evolve together, due to a correlated structural or functional stress at these positions. Co‐evolution has thus been evidenced computationally in multiple proteins or protein domains. Here, we wish to study whether an evolutionary stress is exerted on a sequence alignment across protein domains, i.e., on longer sequence separations than within a single protein domain. JmjC‐containing lysine demethylases were chosen for analysis, as a follow‐up to previous studies; these proteins are important multidomain epigenetic regulators. In these proteins, the JmjC domain is responsible for the demethylase activity, and surrounding domains interact with histones, DNA or partner proteins. This family of enzymes was analyzed at the sequence level, in order to determine whether the sequence of JmjC‐domains was affected by the presence of a neighboring JmjN domain or PHD finger in the protein. Multiple positions within JmjC sequences were shown to have their residue distributions significantly altered by the presence of the second domain. Structural considerations confirmed the relevance of the analysis for JmjN‐JmjC proteins, while among PHD‐JmjC proteins, the length of the linker region could be correlated to the residues observed at the most affected positions. The correlation of domain architecture with residue types at certain positions, as well as that of overall architecture with protein function, is discussed. The present results thus evidence the existence of an across‐domain evolutionary stress in JmjC‐containing demethylases, and provide further insights into the overall domain architecture of JmjC domain‐containing proteins.