Regulation and activity of cytosolic 5'-nucleotidase II. A bifunctional allosteric enzyme of the Haloacid Dehalogenase superfamily involved in cellular metabolism

In many vertebrate tissues, cytosolic 5'-nucleotidase II (cN-II) either hydrolyses or phosphorylates a number of purine (monophosphorylated) nucleosides through a scheme common to the Haloacid Dehalogenase superfamily members. It possesses a pivotal role in purine cellular metabolism and it acts on anti-tumoural and antiviral nucleoside analogues, thus being of potential therapeutic importance. cN-II is Mg2+-dependent, regulated and stabilised by several factors such as allosteric effectors ATP and 2,3-DPG, although these are not directly involved in the reaction stoichiometry. We review herein the experimental knowledge currently available about this remarkable enzymatic activity.     

烟气脱硫石膏对滨海农耕土壤磷素形态组成的影响

为探明不同烟气脱硫石膏施用量对滨海农耕土壤中的全磷、有效磷、无机磷组分等的影响,通过田间试验的方式,分别在试验区土壤中施加0t/hm~2、15t/hm~2、30t/hm~2、45t/hm~2烟气脱硫石膏。研究结果表明:与对照组相比,各处理组的土壤全磷含量无显著差异,而土壤中的有效磷和渗滤液中的可溶性磷含量则随着烟气脱硫石膏施入量的增加而降低;施入烟气脱硫石膏后农耕土壤中无机磷含量显著增加,其中又以磷酸钙盐含量的增加为主,磷酸钙盐中的Ca2-P、Ca_8-P和Ca_10-P含量分别增加了30.8%—68.9%、35.2%—66.3%和7.3%—17.8%。烟气脱硫石膏的施用促进了植物的生长发育,有效磷的降低和无机磷组分中磷酸钙盐的增加并未影响到植物对磷素的吸收。因此,烟气脱硫石膏能有效地固定滨海农耕土壤中的溶解态磷,控制土壤过量磷素向水体迁移,降低附近水体富营养化发生的机率,保障区域水体生态系统环境安全。     

Preliminary results on the photoluminescence and optically stimulated luminescence in Cu‐doped and Ag‐doped ZnB2X4 (B = Li,Na, K: X = Cl,Br) compounds

Photoluminescence, and optically stimulated luminescence in ZnB₂X₄ (B; Li,Na,K: X; Cl,Br) compounds doped with Cu⁺ or Ag⁺ were studied. Double humped emission bands attributable to the activators were observed in all the samples. The observed photoluminescence of Cu⁺ and Ag⁺ could be identified with 3d⁹4s¹?3d¹⁰ and 4d⁹5s¹?5d¹⁰ transitions respectively. The longer wavelength band (400–500 nm range) could be attributed to the Cu⁺ or Ag⁺ ion replacing alkali ion at the octahedral alkali site whereas short wavelength band (340–400 nm range) is attributed to a Cu or Ag ion at tetrahedral zinc site. The short wavelength band was found to be intense compared with long wavelength and gave an indication that most of the Cu or Ag ions prefered a tetrahedral Zn site compared with the octahedral alkali site. All the samples exhibit optically stimulated luminescence (OSL). The sensitivity was found to be lattice dependent. The lowest sensitivity of about 1% compared with Al₂O₃:C was observed in lithium lattices whereas highest the sensitivity of about 290% was observed in the case of Cu‐doped ZnNa₂Br₄.     

饲料中维生素和无机盐对rBmNPV-Bm表达系统植酸酶基因表达活性的影响

【目的】为查明宿主的营养对重组家蚕杆状病毒-家蚕表达系统(rBmNPV-Bm)外源基因表达活性的影响,探寻提高家蚕Bombyx mori生物反应器产率的新途径。【方法】以家蚕BmNPV病毒为表达载体,以人工饲料饲养的5龄家蚕为宿主,以植酸酶基因为报告基因,探讨了家蚕人工饲料中维生素和无机盐对外源基因表达产物活性的影响。【结果】家蚕血淋巴中外源植酸酶的活性随着饲料中维生素和无机盐添加量的不同而发生显著变化。在本试验设区范围内,当维生素C的含量为饲料干物质量的1 %,B族维生素混合物为0.25 %,无机盐混合物为1 %,维生素B6为100 g饲料干物中含2 mg时,血淋巴中植酸酶活性达到最大值。【结论】通过改变家蚕饲料中微量营养成分的含量是提高rBmNPV-Bm表达系统外源基因表达活性的有效途径。     

Efficient proteolysis using a regenerable metal-ion chelate immobilized enzyme reactor supported on organic-inorganic hybrid silica monolith

A metal‐ion chelate immobilized enzyme reactor (IMER) supported on organic–inorganic hybrid silica monolith was developed for rapid digestion of proteins. The monolithic support was in situ prepared in a fused silica capillary via the polycondensation between tetraethoxysilane hydrolytic sol and iminodiacetic acid conjugated glycidoxypropyltrimethoxysilane. After activated by Cu²⁺, trypsin was immobilized onto the monolithic support via metal chelation. Proteolytic capability of such an IMER was evaluated by the digestion of myoglobin and BSA, and the digests were further analyzed by microflow reversed‐phase liquid chromatography with ESI‐MS/MS. Similar sequence coverages of myoglobin and BSA were obtained by IMER, in comparison to those obtained by in‐solution digestion (91 versus 92% for 200 ng myoglobin, and 26 versus 26% for 200 ng BSA). However, the digestion time was shortened from 12 h to 50 s. When the enzymatic activity was decreased after seven runs, the IMER could be easily regenerated by removing Cu²⁺ via EDTA followed by trypsin immobilization with fresh Cu²⁺ introduced, yielding the equal sequence coverage (26% for 200 ng BSA). For ～5 μg rat liver extract, even more proteins were identified with the immobilized trypsin digestion within 150 s in comparison to the in‐solution digestion for 24 h (541 versus 483), demonstrating that the IMER could be a promising tool for efficient and high‐throughput proteome profiling.     

Biogeography of copepods in lakes and ponds of subarctic Québec,Canada

We aimed to determine whether the copepod assemblages in lakes and ponds of northern Québec, Canada, were closer in composition to those found in southern Québec, or to those reported from the subarctic and arctic. Six calanoid and five cyclopoid species were identified from 37 ponds and lakes located in the region between 55° N and 59° N. Species diversity was generally low, ranging from 0 to 4 species per lake. Dominant species were Leptodiaptomus minutus and Acanthocyclops vernalis. The species assemblages showed high affinity with those found in forested regions of southern Québec. Exceptions were Leptodiaptomus tyrrelli, previously only recorded west of Hudson Bay, and Hesperodiaptomus arcticus, hitherto recorded north of 58° N. Relationships between the lakes, species, and environmental variables were explored using multivariate analysis. Lakes situated along the coast and on two offshore islands clustered together and were characterised by higher conductivity and pH than those lakes located further inland. Leptodiaptomus tyrrelli was common in these coastal lakes. Canonical correspondence analysis revealed statistically significant relationships between copepod distributions and conductivity, dissolved organic carbon and pH. These three variables accounted for 70% of the variation in the species' distribution.     

Oligomerization of H-pyrophosphatase and its structural and functional consequences

The H⁺-pyrophosphatase (H⁺-PPase) consists of a single polypeptide, containing 16 or 17 transmembrane domains. To determine the higher order oligomeric state of Streptomyces coelicolor H⁺-PPase, we constructed a series of cysteine substitution mutants and expressed them in Escherichia coli. Firstly, we analyzed the formation of disulfide bonds, promoted by copper, in mutants with single cysteine substitutions. 28 of 39 mutants formed disulfide bonds, including S545C, a substitution at the periplasmic side. The formation of intermolecular disulfide bonds suppressed the enzyme activity of several, where the substituted residues were located in the cytosol. Creating disulfide links in the cytosol may interfere with the enzyme's catalytic function. Secondly, we prepared double mutants by introducing second cysteine substitutions into the S545C mutant. These double-cysteine mutants produced cross-linked complexes, estimated to be at least tetramers and possibly hexamers. Thirdly, we co-expressed epitope-tagged, wild type, and inactive mutant H⁺-PPases in E. coli and confirmed the formation of oligomers by co-purifying one subunit using the epitope tag used to label the other. The enzyme activity of these oligomers was markedly suppressed. We propose that H⁺-PPase is present as an oligomer made up of at least two or three sets of dimers.     

Measurement of the inorganic pyrophosphate in tissues of Pisum sativum L.

Purified pyrophosphate: fructose 6-phosphate 1-phosphotransferase (EC 2.7.1.90) was used to measure the inorganic pyrophosphate in unfractionated extracts of tissues of Pisum sativum L. The fructose 1,6-bisphosphate produced by the above enzyme was measured by coupling to NADH oxidation via aldolase (EC 4.1.2.13), triosephosphate isomerase (EC 5.3.1.1) and glycerol-3-phosphate dehydrogenase (EC 1.1.1.8). Amounts of pyrophosphate as low as 1 nmol could be measured. The contents of pyrophosphate in the developing embryo of pea, and in the apical 2 cm of the roots, were appreciable; 9.4 and 8.9 nmol g^-1 fresh weight, respectively. The possibility that pyrophosphate acts in vivo as an energy source for pyrophosphate: fructose 6-phosphate 1-phosphotransferase and for UDPglucose pyrophosphorylase (EC 2.7.7.9) is considered.     

有机无机肥混施对水稻土CH4排放的影响

通过网室水稻盆栽试验,探讨几种有机无机肥混施方式对CH₄排放的影响.结果表明,水稻土CH₄排放有明显的逐日变化,在水稻分蘖末期和抽穗前期出现CH₄排放高峰.不同有机无机肥混施方式对CH₄排放的影响明显,最高和最低的CH₄排放量分别为8.12和5.72g·m^-2,两者相差42.0%.粒状有机无机复肥的CH₄排放量高,但稻谷产量也高,因而其单位稻谷产量的CH₄排放量反而较低.