河川沙塘鳢胚胎、仔鱼发育过程中蛋白质含量及氨基酸库的变化

采用生物化学方法测定和分析了河川沙塘鳢(Odontobutis potamophila)胚胎、仔鱼发育过程中蛋白质含量及氨基酸的组成和含量。结果显示,从受精卵开始到孵出后8 d饥饿仔鱼的整个胚胎和仔鱼发育过程中,其总蛋白和总氨基酸含量均呈下降趋势;总的必需氨基酸含量和总的非必需氨基酸含量也一样都呈现出下降趋势;平均含量最高的氨基酸是亮氨酸Leu、赖氨酸Lys、谷氨酸Glu和天冬氨酸Asp。与总氨基酸库相反,游离氨基酸库却呈现出随着发育的进行而不断增长的趋势,这种增长趋势也反映在游离的必需氨基酸和游离的非必需氨基酸含量的变化上,并且平均含量最高的游离氨基酸是赖氨酸Lys、亮氨酸Leu、脯氨酸Pro和谷氨酸Glu。整个发育过程中,游离氨基酸的含量仅占总氨基酸库的很小一部分(在受精卵,仅占0.16%)。由于总蛋白和总氨基酸含量在发育过程中均呈下降趋势,而游离氨基酸含量呈增长趋势,由此可知,在整个胚胎发生过程中,其卵黄蛋白的水解速度大于胚体同化和异化作用对氨基酸的消耗速度。     

Branched Chain Amino Acid Metabolism in the Biosynthesis of Lycopersicon pennellii Glucose Esters

Lycopersicon pennellii Corr. (D'Arcy) an insect-resistant, wild tomato possesses high densities of glandular trichomes which exude a mixture of 2,3,4-tri-O-acylated glucose esters that function as a physical impediment and feeding deterrent to small arthropod pests. The acyl moieties are branched C₄ and C₅ acids, and branched and straight chain C₁₀, C₁₁, and C₁₂ acids. The structure of the branched acyl constituents suggests that the branched chain amino acid biosynthetic pathway participates in their biosynthesis. [¹⁴C]Valine and deuterated branched chain amino acids (and their oxo-acid derivatives) were incorporated into branched C₄ and C₅ acid groups of glucose esters by a process of transamination, oxidative decarboxylation and subsequent acylation. C₄ and C₅ branched acids were elongated by two carbon units to produce the branched C₁₀-C₁₂ groups. Norvaline, norleucine, allylglycine, and methionine also were processed into acyl moieties and secreted from the trichomes as glucose esters. Changes in the acyl composition of the glucose esters following sulfonylurea herbicide administration support the participation of acetohydroxyacid synthetase and the other enzymes of branched amino acid biosynthesis in the production of glucose esters.     

Carbohydrate and Amino Acid Metabolism in the Ectomycorrhizal Ascomycete Sphaerosporella brunnea during Glucose Utilization : A C NMR Study

Nuclear magnetic resonance spectroscopy was utilized to study the metabolism of [1-¹³C]glucose in mycelia of the ectomycorrhizal ascomycete Sphaerosporella brunnea. The main purpose was to assess the biochemical pathways for the assimilation of glucose and to identify the compounds accumulated during glucose assimilation. The majority of the ¹³C label was incorporated into mannitol, while glycogen, trehalose and free amino acids were labeled to a much lesser extent. The high enrichment of the C1/C6 position of mannitol indicated that the polyol was formed via a direct route from absorbed glucose. Randomization of the ¹³C label was observed to occur in glucose and trehalose leading to the accumulation of [1,6-¹³C]trehalose and [1,6-¹³C]glucose. This suggests that the majority of the glucose carbon used to form trehalose was cycled through the metabolically active mannitol pool. The proportion of label entering the free amino acids represented 38% of the soluble ¹³C after 6 hours of continuous glucose labeling. Therefore, amino acid biosynthesis is an important sink of assimilated carbon. Carbon-13 was incorporated into [3-¹³C]alanine and [2-¹³C]-, [3-¹³C]-, and [4-¹³C]glutamate and glutamine. From the analysis of the intramolecular ¹³C enrichment of these amino acids, it is concluded that [3-¹³C]pyruvate, arising from [1-¹³C]glucose catabolism, was used by alanine aminotransferase, pyruvate dehydrogenase, and pyruvate carboxylase (or phosphoenolpyruvate carboxykinase). Intramolecular ¹³C labeling patterns of glutamate and glutamine were similar and are consistent with the operation of the Krebs cycle. There is strong evidence for (a) randomization of the label on C2 and C3 positions of oxaloacetate via malate dehydrogenase and fumarase, and (b) the dual biosynthetic and respiratory role of the citrate synthase, aconitase, and isocitrate dehydrogenase reactions. The high flux of carbon through the carboxylation (presumably pyruvate carboxylase) step indicates that CO₂ fixation is an important component of the carbon metabolism in S. brunnea, and it is likely that this anaplerotic role is particularly prevalent during NH₄⁺ assimilation. The most relevant information resulting from this investigation is (a) the occurrence of the mannitol cycle, (b) a large part of the trehalose pool is synthesized after the cycling of glucose-carbon through the mannitol cycle, and (c) pyruvate (or phosphoenolpyruvate) carboxylation plays an important role in the primary metabolism of glucose-fed mycelia.     

Amino Acid Metabolism in Rat Hippocampus During the Period of Brain Growth Spurt

We studied protein synthesis, lipid synthesis and CO₂ production by oxidation of glycine, alanine and leucine by slices of rat hippocampus during the period of brain growth spurt. The metabolism of the three amino acids decreased with the age of the animals, A major reduction was observed in protein synthesis, which was 4 times higher at 7 days of age than at 21 days of age for all amino acids studied. Glycine oxidation to CO₂ was twice as high as alanine oxidation and ten times higher than leucine oxidation. The major pathway of leucine utilization was incorporation into proteins. Glycine was the amino acid that had the highest metabolic rate.     

Amino Acid Availability Regulates the Effect of Hyperinsulinemia on Skin Protein Metabolism in Pigs

The effects of amino acid supply and insulin infusion on skin protein kinetics (fractional synthesis rate (FSR), fractional breakdown rate (FBR), and net balance (NB)) in pigs were investigated. Four-month-old pigs were divided into four groups as follows: control, insulin (INS), amino acid (AA), and INS + AA groups based on the nutritional and hormonal conditions. l-[ring-¹³C₆]Phenylalanine was infused. FBR was estimated from the enrichment ratio of arterial phenylalanine to intracellular free phenylalanine. Plasma INS was increased (p < 0.05) in the INS and INS + AA groups. Plasma glucose was maintained by infusion of glucose in the groups receiving INS. The interventions did not change the NB of skin protein. However, the interventions affected the FSR and FBR differently. An infusion of INS significantly increased both FSR and FBR, although AA infusion did not. When an AA infusion was added to the infusion of insulin (INS + AA group), FSR and FBR were both lower when compared with the INS group. Our data demonstrate that in anesthetized pigs INS infusion did not exert an anabolic effect, but rather it increased AA cycling into and out of skin protein. Because co-infusion of AAs with INS ameliorated this effect, it is likely that the increased AA cycling during INS infusion was related to AA supply. Although protein kinetics were affected by both INS and AAs, none of the interventions affected the skin protein deposition. Thus, skin protein content is closely regulated under normal circumstances and is not subject to transient changes in AAs or hormonal concentrations.     

非编码RNA与肝脏糖脂代谢调控

随着经济的迅速发展、人们饮食习惯的改变和身体活动的减少,糖尿病成为了现代社会的非传染性的流行病,给家庭和社会造成了极大的危害和经济负担。其中以全身性胰岛素抵抗及胰岛功能衰竭为主要发病特征的2型糖尿病(type 2 diabetes,T2D)已在世界范围引起广泛关注。T2D的发生发展涉及许多组织及糖代谢的各个环节,遗传因素和环境因素共同引起的糖脂代谢通路任一环节的失调均可导致T2D的发生。近年来,包括microRNA(miRNA)及长非编码RNA(LncRNA)在内的非编码RNA(ncRNA)的发现及其在人体生理和病理生理过程中的重要调控作用不断被揭示,为进一步了解T2D的发病机制注入了新理念和信息。miRNAs及LncRNAs的表达具有组织特异性,其表达水平的异常通常与疾病相关。本文主要对miRNAs和LncRNAs在肝脏糖脂代谢调控及T2D的发生发展中的作用及机制的最新研究进展作简要综述。     

Glucose Metabolism and Kinetics of Phosphorus Removal by the Fermentative Bacterium Microlunatus phosphovorus

Phosphorus and carbon metabolism in Microlunatus phosphovorus was investigated by using a batch reactor to study the kinetics of uptake and release of extracellular compounds, in combination with ³¹P and ¹³C nuclear magnetic resonance (NMR) to characterize intracellular pools and to trace the fate of carbon substrates through the anaerobic and aerobic cycles. The organism was subjected to repetitive anaerobic and aerobic cycles to induce phosphorus release and uptake in a sequencial batch reactor; an ultrafiltration membrane module was required since cell suspensions did not sediment. M. phosphovorus fermented glucose to acetate via an Embden-Meyerhof pathway but was unable to grow under anaerobic conditions. A remarkable time shift was observed between the uptake of glucose and excretion of acetate, resulting in an intracellular accumulation of acetate. The acetate produced was oxidized in the subsequent aerobic stage. Very high phosphorus release and uptake rates were measured, 3.34 mmol g of cell⁻¹ h⁻¹ and 1.56 mmol g of cell⁻¹ h⁻¹, respectively, values only comparable with those determined in activated sludge. In the aerobic period, growth was strictly dependent on the availability of external phosphate. Natural abundance ¹³C NMR showed the presence of reserves of glutamate and trehalose in cell suspensions. Unexpectedly, [1-¹³C]glucose was not significantly channeled to the synthesis of internal reserves in the anaerobic phase, and acetate was not during the aerobic stage, although the glutamate pool became labeled via the exchange with intermediates of the tricarboxylic acid cycle at the level of glutamate dehydrogenase. The intracellular pool of glutamate increased under anaerobic conditions and decreased during the aerobic period. The contribution of M. phosphovorus for phosphorus removal in wastewater treatment plants is discussed on the basis of the metabolic features disclosed by this study.     

Involvement of Calcium and Calmodulin in Protein and Amino Acid Metabolism in Rice Roots under Anoxia

The pretreatment of rice roots for 1 h in aerobic conditionswith the Ca²⁺-channel blockers ruthenium red (RR) and verapamiland the calmodulin (CaM) antagonists N-(6-aminohexyl)-5-chloro-1-naphtylenesulfonamide(W-7) and trifluoperazine, induced during 3 h of anoxia: (i)inhibition of amino acid and     

脂联素对肝脏糖脂代谢调节作用的研究进展

脂联素因其具有抗糖尿病作用而备受关注,它能控制血糖,并且在肝脏、脂肪和胰腺中能影响脂质代谢。脂联素通过刺激脂肪细胞,对抗炎症、控制脂质过氧化和脂肪分解的速率来调控脂质流入非脂肪组织。肝脏是脂联素发挥作用的重要靶器官;在肝脏中,脂联素与脂联素受体1、2或T-钙黏着蛋白结合,激活下游的AMPK、APPL1、神经酰胺酶等发挥其调节作用。我们总结了脂联素改善肝脏胰岛素敏感性和糖脂代谢的相关机制。     

Modification of the Induction of Photosynthesis in Wheat by Glyphosate, an Inhibitor of Amino Acid Metabolism

Ireland, C. R., Pcrcival, M. P. and Baker, N. R. 1986. Modificationof the induction of photosynthesis in wheat by glyphosate, aninhibitor of amino acid metabolism.—J. exp. Bot. 37: 299–308 The effect of the herbicide glyphosate, an inhibitor of aminoacid metabolism, upon the kinetics of induction of photosynthesiswas investigated in wheat leaves. After treatment with a commercialpreparation of glyphosate the induction period for both photosyntheticcarbon assimilation and chlorophyll fluorescence emission wasincreased approximately by 50% after 2 d and by 100%) after5 d. The quenching of chlorophyll fluorescence became a slowerprocess and the S-M transient was diminished. In the steady-stateneither the rate of carbon fixation nor the level of fluorescenceemission were altered. The changes in induction kinetics precededthe occurrence of visible leaf damage. The fluorescence emissionwas analysed by a technique which estimates the redox stateof the PS2 primary electron acceptor, Q, and the photochemicaland non-photochemical components of fluorescence quenching.Glyphosate produced no fundamental change in the relationshipbetween the steady-state values of these parameters until thetime when extensive leaf damage occurred. Thus there was nodirect effect of glyphosate on the capacity for light captureor on the rate of photosynthetic electron transport. This wasconfirmed by the absence of any effect on the quantum efficiencyof oxygen evolution. The slower rate of fluorescence quenchingduring induction is attributed to a slower generation of thetrans-thylakoid pH. It is proposed that glyphosate affects photosyntheticinduction kinetics by an indirect modification of carbon metabolismwhich limits photosynthetic rate during this phase. The possibilityof screening for rapid effects of non-photosynthetic herbicides,such as glyphosate, by monitoring the kinetics of photosyntheticinduction, is raised. Key words: Photosynthesis, induction, glyphosate (N-phosphonomethyl-glycine), wheat (Triticum aestivum L.)     

Protein and Nucleic Acid Metabolism in Fruits: I. Studies of Amino Acid Incorporation During the Climacteric Rise in Respiration of the Avocado

Incorporation of (14)C L-valine and (14)C L-leucine into protein of tissue slices of the avocado fruit was relatively high during the early stages of the climacteric rise, declined sharply thereafter, and was virtually absent at the peak. The incorporation of amino acids in the preclimacteric stage was markedly lower than during the early stage of the respiratory rise. By following incorporation in relation to uptake at several concentrations it was established that the results were not a reflection of endogenous dilution.Puromycin was effective as an inhibitor of incorporation but not of oxygen uptake. When respiration was at its maximum there was no protein synthesis. It was concluded, therefore, that the respiratory upsurge characteristic of the climacteric was not related directly to protein synthesis.     

Interrelationship of Growth and Plasma Amino Acid Concentration in the Chick Fed Casein Diets as Influenced by Dietary Protein and Amino Acid Levels

Chicks fed a 20% casein-sucrose diet showed severe growth depression, but dietary supplementation with 0.7% arginine-HCl, 0.35% glycine and 0.35% dl-methionine improved the growth rate to almost that of chicks fed a practical diet. Chicks fed high protein diets containing 64% casein showed normal growth, irrespective of the supplementation with such amino acids. Plasma arginine concentration of growth-retarded chicks was significantly lower than that of normal chicks. Plasma threonine/arginine ratio was negatively correlated with the growth rate of the chicks, the ratio of normal chicks being 3 ~ 4 whereas that of growth-retarded chicks was about 24. No lysine-arginine antagonism occurred-under high casein feeding.     

CcpA-Mediated Enhancement of Sugar and Amino Acid Metabolism in Lysinibacillus sphaericus by NMR-Based Metabolomics

Lysinibacillus sphaericus is a bacterium incapable of metabolizing sugars with the sole exception of N-acetylglucosamine. To unravel the regulatory role of catabolite control protein A (CcpA) in the sugar metabolism of L. sphaericus, a ccpA deficient mutant was constructed by homologous recombination. The mutant showed growth deficiency and a low efficiency of carbon and energy utilization. NMR spectroscopy in combination with multivariate data analysis revealed that the metabolome of L. sphaericus was dominated by 25 metabolites mainly including amino acids, carbohydrate derivatives and organic acids, and that the mutation of the ccpA gene caused significant reduction of leucine, valine, alanine, threonine, glutamate, lysine, d-ornithine, tyrosine, uridine 5'-diphospho-N-acetlyglucosamine formate, fumarate, phenylalanine, aspartate, asparagine, and acetate but elevation of ribose-5-phosphate, and uracil. Furthermore, the networks of CcpA-mediated regulation based on the metabolome were constructed by arrangement of significantly decreasing or increasing metabolites. The network map suggests CcpA regulates and promotes sugar and amino acid metabolism of L. sphaericus.