Structure of the GDP domain of EF-Tu and location of the amino acids homologous to ras oncogene proteins

A 2.7 angstrom resolution x-ray diffraction analysis of a trypsin-modified form of the Escherichia coli elongation factor Tu reveals that the GDP-binding domain has a structure similar to that of other nucleotide-binding proteins. The GDP ligand is located at the COOH-terminal end of the beta sheet and is linked to the protein via a Mg2+ ion salt bridge. The location of the guanine ring is unusual; the purine ring is located on the outer edge of the domain, not deep within a hydrophobic pocket. The amino acids from Pro10 to Arg44 and from Gly59 to Glu190 have been assigned to the electron density with computer graphic techniques, and the resulting model is consistent with all known biochemical data. An analysis of the structure reveals that four regions of the amino acid sequence that are homologous with the family of ras oncogene proteins, termed p21, are located in the vicinity of the GDP-binding site, and most of the invariant amino acids shared by the proteins interact directly with the GDP ligand.     

Identification of a specialized adenylyl cyclase that may mediate odorant detection

The mammalian olfactory system may transduce odorant information via a G protein-mediated adenosine 3',5'-monophosphate (cAMP) cascade. A newly discovered adenylyl cyclase, termed type III, has been cloned, and its expression was localized to olfactory neurons. The type III protein resides in the sensory neuronal cilia, which project into the nasal lumen and are accessible to airborne odorants. The enzymatic activity of the type III adenylyl cyclase appears to differ from nonsensory cyclases. The large difference seen between basal and stimulated activity for the type III enzyme could allow considerable modulation of the intracellular cAMP concentration. This property may represent one mechanism of achieving sensitivity in odorant perception.     

mTORC1 senses lysosomal amino acids through an inside-out mechanism that requires the vacuolar H(+)-ATPase

The mTOR complex 1 (mTORC1) protein kinase is a master growth regulator that is stimulated by amino acids. Amino acids activate the Rag guanosine triphosphatases (GTPases), which promote the translocation of mTORC1 to the lysosomal surface, the site of mTORC1 activation. We found that the vacuolar H(+)-adenosine triphosphatase ATPase (v-ATPase) is necessary for amino acids to activate mTORC1. The v-ATPase engages in extensive amino acid-sensitive interactions with the Ragulator, a scaffolding complex that anchors the Rag GTPases to the lysosome. In a cell-free system, ATP hydrolysis by the v-ATPase was necessary for amino acids to regulate the v-ATPase-Ragulator interaction and promote mTORC1 translocation. Results obtained in vitro and in human cells suggest that amino acid signaling begins within the lysosomal lumen. These results identify the v-ATPase as a component of the mTOR pathway and delineate a lysosome-associated machinery for amino acid sensing.     

The Rag GTPases bind raptor and mediate amino acid signaling to mTORC1

The multiprotein mTORC1 protein kinase complex is the central component of a pathway that promotes growth in response to insulin, energy levels, and amino acids and is deregulated in common cancers. We find that the Rag proteins--a family of four related small guanosine triphosphatases (GTPases)--interact with mTORC1 in an amino acid-sensitive manner and are necessary for the activation of the mTORC1 pathway by amino acids. A Rag mutant that is constitutively bound to guanosine triphosphate interacted strongly with mTORC1, and its expression within cells made the mTORC1 pathway resistant to amino acid deprivation. Conversely, expression of a guanosine diphosphate-bound Rag mutant prevented stimulation of mTORC1 by amino acids. The Rag proteins do not directly stimulate the kinase activity of mTORC1, but, like amino acids, promote the intracellular localization of mTOR to a compartment that also contains its activator Rheb.     

利用特征氨基酸组分分析与显微观察鉴别掺假鱼粉

通过显微镜检观察,发现两类掺假鱼粉中分别含有海泥颗粒或羽毛碎片等杂质.氨基酸分析结果表明,掺假鱼粉的氨基酸总量和蛋氨酸、赖氨酸、苏氨酸、色氨酸等必需氨基酸含量大幅度降低,造成氨基酸组分不平衡;掺入海泥的鱼粉中芳香族氨基酸--酪氨酸含量不到合格鱼粉正常值的1/4,而掺入羽毛粉的鱼粉,丝氨酸、胱氨酸、脯氨酸等含量反而比优质鱼粉的正常含量还要高.分析结果还表明,优质或合格鱼粉中检出丰富的牛磺酸(637.4～746.8 mg·hg-1),而掺假鱼粉中牛磺酸含量只有33.27～43.76 mg·kg-1,不到正常值的1/10,可以作为特征性指标成分进行鱼粉是否掺假的检测分析.采用氨基酸组分分析技术结合显微镜观察可以准确鉴别掺假鱼粉.     

Glutamatergic and dopaminergic neurons mediate anxiogenic and anxiolytic effects of CRHR1

The corticotropin-releasing hormone receptor 1 (CRHR1) critically controls behavioral adaptation to stress and is causally linked to emotional disorders. Using neurochemical and genetic tools, we determined that CRHR1 is expressed in forebrain glutamatergic and γ-aminobutyric acid-containing (GABAergic) neurons as well as in midbrain dopaminergic neurons. Via specific CRHR1 deletions in glutamatergic, GABAergic, dopaminergic, and serotonergic cells, we found that the lack of CRHR1 in forebrain glutamatergic circuits reduces anxiety and impairs neurotransmission in the amygdala and hippocampus. Selective deletion of CRHR1 in midbrain dopaminergic neurons increases anxiety-like behavior and reduces dopamine release in the prefrontal cortex. These results define a bidirectional model for the role of CRHR1 in anxiety and suggest that an imbalance between CRHR1-controlled anxiogenic glutamatergic and anxiolytic dopaminergic systems might lead to emotional disorders.     

大豆异黄酮对大鼠小肠上皮细胞生长及葡萄糖和氨基酸吸收的影响

分离培养了SD大鼠的小肠上皮细胞, 用3H TdR掺入法分别检测了 0、0 1、1、10、100和 1 000ng·mL-1 2种大豆异黄酮 (大豆黄酮和染料木素) 对该细胞生长的影响, 并测定了细胞碱性磷酸酶活性的变化。同时, 利用翻转的离体小肠囊研究了 0、0 1、0 5、1和 5μg·mL-1大豆异黄酮对小肠葡萄糖与氨基酸吸收的影响, 并检测了小肠黏膜总ATP酶活性的变化。结果显示: 100ng·mL-1的大豆异黄酮 (大豆黄酮、染料木素 ) 能够提高该细胞3H TdR的掺入量和碱性磷酸酶的活性, 提示大豆异黄酮可能有促小肠细胞生长的活性; 0 1和 0 5ng·mL-1的大豆黄酮和 0 5和 1 0ng·mL-1的染料木素对离体小肠葡萄糖的吸收具有促进作用, 而对赖氨酸与色氨酸的吸收以及小肠黏膜总ATP酶的活性没有明显影响。     

不同品种精白米必需氨基酸、总氨基酸和蛋白质含量的相关分析

测定了28个精白米样品的必需氨基酸、总氨基酸、蛋白质含量,分析了不同精白样品各成分的相关性。结果表明,28个精白米样品蛋白质含量与总氨基酸含量、蛋白质含量与必需氨基酸含量、总氨基酸含量与必需氨基酸含量的相关系数分别达到0.940、0.950、0.995,均呈极显著正相关。精白米主要营养成分蛋白质的营养价值主要在于氨基酸含量的高低,必需氨基酸含量可作为判定稻米营养价值的重要指标。     

Inhibition of GTPase activating protein stimulation of Ras-p21 GTPase by the Krev-1 gene product

Krev-1 is known to suppress transformation by ras. However, the mechanism of the suppression is unclear. The protein product of Krev-1, Rap1A-p21, is identical to Ras-p21 proteins in the region where interaction with guanosine triphosphatase (GTPase) activating protein (GAP) is believed to occur. Therefore, the ability of GAP to interact with Rap1A-p21 was tested. Rap1A-p21 was not activated by GAP but bound tightly to GAP and was an effective competitive inhibitor of GAP-mediated Ras-GTPase activity. Binding of GAP to Rap1A-p21 was strictly guanosine triphosphate (GTP)-dependent. The ability of Rap1A-p21 to bind tightly to GAP may account for Krev-1 suppression of transformation by ras. This may occur by preventing interaction of GAP with Ras-p21 or with other cellular proteins necessary for GAP-mediated Ras GTPase activity.     

氨基酸胁迫对小麦种子幼苗生长和体内游离氨基酸水平的影响

摘 要：用赖氨酸、苏氨酸两种氨基酸单因子胁迫和两因子协同胁迫不同蛋白质含量小麦品种种子的发芽幼苗,发现5 mmol/l浓度可严重抑制根系生长,但对叶生长抑制较轻;两种氨基酸具有协同抑制作用。相比较,高蛋白含量基因型品种比低蛋白含量基因型品种可忍耐较高浓度氨基酸胁迫。HPLC方法测定胁迫植株体内游离氨基酸含量,发现胁迫氨基酸有比对照高3～10倍积累,同族氨基酸有比对照高2～3倍积累,相关氨基酸也有1～2倍积累。     

Dense packing and symmetry in small clusters of microspheres

When small numbers of colloidal microspheres are attached to the surfaces of liquid emulsion droplets, removing fluid from the droplets leads to packings of spheres that minimize the second moment of the mass distribution. The structures of the packings range from sphere doublets, triangles, and tetrahedra to exotic polyhedra not found in infinite lattice packings, molecules, or minimum-potential energy clusters. The emulsion system presents a route to produce new colloidal structures and a means to study how different physical constraints affect symmetry in small parcels of matter.     

Wheat straw biochar amendment suppresses tomato bacterial wilt caused by Ralstonia solanacearum: Potential effects of rhizosphere organic acids and amino acids

Complex interactions based on host plant, rhizosphere microorganisms and soil microenvironment are presumed to be responsible for the suppressive properties of biochar against soil-borne diseases, although the underlying mechanisms are not well understood. This study is designed to evaluate the efficacy of biochar amendment for controlling tomato bacterial wilt caused by Ralstonia solanacearum, and to explore the interactions between biochar-induced changes in rhizosphere compound composition, the pathogen and tomato growth. The results showed that biochar amendment decreased disease incidence by 61–78% and simultaneously improved plant growth. The positive ‘biochar effect' could be associated with enhanced microbial activity and alterations in the rhizosphere organic acid and amino acid composition. Specifically, elevated rhizosphere citric acid and lysine, but reduced salicylic acid, were induced by biochar which improved microbial activity and rendered the rhizosphere unsuitable for the development of R. solanacearum. In addition, nutrients which were either made more available by the stimulated microbial activity or supplied by the biochar could improve plant vigor and potentially enhance tomato resistance to diseases. Our findings highlight that biochar's ability to control tomato bacterial wilt could be associated with the alteration of the rhizosphere organic acid and amino acid composition, however, further research is required to verify these ‘biochar effects' in field conditions.     

猪链球菌9型噬菌体裂解酶Ply5218最小功能域及关键氨基酸位点的鉴定

为了鉴定猪链球菌9型噬菌体裂解酶Ply5218的最小活性功能域及关键氨基酸位点,利用PCR技术对全长裂解酶Ply5218进行截短并对可能与活性相关的关键氨基酸位点进行定点突变,研究截短与突变后各个蛋白的活性并与全长蛋白活性对比。结果显示,截短片段Ply5218_(1-147)可在平板上形成裂菌圈,仍保持裂菌活性,而比该片段更短的截短片段则失去裂菌活性,推测Ply5218_(1-147)为Ply5218的最小活性相关功能域;第8、58、136和142位点的氨基酸突变后,裂菌活性部分降低,第34和144位点的氨基酸突变后,则彻底失去裂菌活性。研究结果为深入揭示Ply5218的裂菌机制和进一步改造裂解酶提供了基础数据。     

氨基酸螯合稀土微肥(大丰收)的应用效果

氨基酸螯合稀土微肥(大丰收)由氨基酸、稀土、微量元素等组成,对其在多种农作物上进行了试验、示范,结果表明,对水稻、西瓜、黄瓜、辣椒、番茄和茄子等多种作物均表现出显著的增产效果,同时还能改善农作物的品质。     

LAAT-1 is the lysosomal lysine/arginine transporter that maintains amino acid homeostasis

Defective catabolite export from lysosomes results in lysosomal storage diseases in humans. Mutations in the cystine transporter gene CTNS cause cystinosis, but other lysosomal amino acid transporters are poorly characterized at the molecular level. Here, we identified the Caenorhabditis elegans lysosomal lysine/arginine transporter LAAT-1. Loss of laat-1 caused accumulation of lysine and arginine in enlarged, degradation-defective lysosomes. In mutants of ctns-1 (C. elegans homolog of CTNS), LAAT-1 was required to reduce lysosomal cystine levels and suppress lysosome enlargement by cysteamine, a drug that alleviates cystinosis by converting cystine to a lysine analog. LAAT-1 also maintained availability of cytosolic lysine/arginine during embryogenesis. Thus, LAAT-1 is the lysosomal lysine/arginine transporter, which suggests a molecular explanation for how cysteamine alleviates a lysosomal storage disease.     

氨基酸平衡性评价指标的比较

为了比较用氨基酸比值系数分(SRC)、氨基酸模式模糊贴近度(FNAAP)评价蛋白质营养价值的合理性,并评估用蛋白质必需氨基酸总含量比值(RTEAA)评价蛋白质营养价值的合理性,用66组文献数据分析了SRC、FNAAP、RTEAA与蛋白质生物价(BV)的相关关系.结果表明,RTEAA与BV极显著相关(ryi.=0.511 6,P<0.001),SRC与BV显著相关(ryi.=0.462 6,P <0.05),FNAAP与BV相关性不显著(P >0.05),由此推断,用SRC评价蛋白质营养价值比用FNAAP合理;RTEAA是评价蛋白质营养价值的合理指标.建议以氨基酸平衡度(degree of essential amino acid balance,DEAAB)这一统计学和生物学含义更为明确的概念替代氨基酸比值系数分(SRC),改用公式DEAAB=1-CVAASs计算氨基酸平衡度,式中CVAASs为氨基酸分(amino acid scores,AASs)的变异系数.     

Synthesis of amino acids by the heating of formaldehyde and ammonia

The heating of formaldehyde and ammonia yields a product that, on hydrolysis, is converted into seven amino acids: aspartic acid, glutamic acid, serine, proline, valine, glycine, and alanine. Glycine is the predominant amino acid. Inasmuch as formaldehyde and ammonia have been identified as compounds in galactic clouds, these experimental results are interpreted in a cosmochemical and geochemical context.     

典型小河流溶解态氨基酸的分布与组成研究

水体中溶解性氨基酸(DAA)是溶解性有机物(DOM)的重要组成部分。为探讨小型河流中DAA的环境意义,于2015年3月(平水期)和6月(丰水期)在西安沣河流域的干流及主要支流采集水样,分析了溶解态氨基酸总量和组成的时空分布特征;并结合氨基酸降解指数(DI)和氨基酸占有机碳的比例(DAA%DOC),探讨了该区域溶解态氨基酸和溶解有机物的来源和迁移转化情况。结果表明:沣河DAA主要来自于外源,丰水期的DAA新鲜程度高于平水期;但由于受到大量降解程度高的有机物稀释,DOM总体的生物有效性降低;由此可见小型河流生产能力弱,主要是DAA的降解场所,丰水期水土流失是DOM进入小型河流的重要途径。由于受水库蓄水影响,支流潏河DAA和DOM的变化特征与沣河干流有显著差异。