Partition of proteins in aqueous two-phase systems containing charged dextran or hydrophobic PEG

Summary The electrochemical effect of a charged dextran derivative and the hydrophobic effect of hydrophobic chain PEG derivative on partitioning of six types of proteins in PEG/dextran aqueous two-phase systems were investigated- When 1. 6%(w/w)DEAE-dextran was present in the system,the partition coefficient decreased quickly with increasing pH value;when 0. 4% (w/w)PEG pentadecanoic acid ester was present in the system, the partition coefficient of protein with strong hydrophobicity was greatly increased. The experimental results show that the influence of hydrocarbon chain PEG derivative on partition coefficient is closely related to the hydrophobicity of proteins.     

谷类作物β-葡聚糖合成酶基因家族的研究进展

Beta-葡聚糖是由β-(1,3)和β-(1,4)糖苷键连接的非纤维素多糖,主要分布在谷类作物籽粒胚乳及糊粉层中,在高尔基体合成,经由囊泡运输到质膜,最终在细胞壁上沉积。通过增加胆汁酸排泄,延迟葡萄糖吸收,β-葡聚糖可有效降低胆固醇及血糖水平。Beta-葡聚糖合成酶基因家族成员最早在水稻(Oryza sativa)中得到鉴定,后在其他作物中陆续被发现。该基因家族包括3个主要成员：CslF、CslH和CslJ亚基因家族,起源于不同分支,经过趋同演化,执行合成β-葡聚糖的功能。Beta-葡聚糖基因家族成员均受到负选择压力,演化过程中序列高度保守。CslF亚家族基因成员相对较多,常在染色体上形成基因簇,CslF6是介导β-葡聚糖合成的主效基因。CslF亚家族在叶基部等幼嫩组织中表达水平相对较高,且明显受到光照强度的影响;CslH和CslJ亚家族成员较少,其中CslH亚家族在叶尖等成熟组织中的表达水平高,而CslJ亚家族在籽粒中有较高的表达水平。该文综述了β-葡聚糖合成酶基因家族成员的系统发育关系、表达模式,β-葡聚糖合成酶的亚细胞定位,以及作物中的定向育种研究进展,提出β-葡聚糖合成酶基因家...     

The Ethanol Extract of Syringa oblata Heartwood,a Mongolian Folk Medicine Containing Major Lignans,Exerts Analgesic and Sedative Effects on Mice

The heartwood of Syringa oblata Lindl. (SO) is one of Mongolian folk medicines to treat insomnia and pain, while its pharmacological evaluation and underlying mechanism remain unclear. In this study, the sedative effect of ethanol extract of SO (ESO) was evaluated with the locomotor activity test and the threshold dose of pentobarbital sodium-induced sleep test in mice, and the hot plate test, acetic acid-induced writhing test, and formalin test in mice were used to evaluate its analgesic effect. The underlying mechanism of ESO analgesia was explored by RT-PCR and western blot analysis, which is associated with the regulation of the NF-κB signaling pathway. Besides, the main constituents of ESO were characterized by LC/MS data analysis and comparison with isolated pure compounds. The current findings brought evidence for clinical application and further pharmacological and phytochemical studies on SO.     

Structural insight into chitin perception by chitin elicitor receptor kinase 1 of Oryza sativa

Plants have developed innate immune systems to fight against pathogenic fungi by monitoring pathogenic signals known as pathogen-associated molecular patterns (PAMP) and have established endo symbiosis with arbuscular mycorrhizal (AM) fungi through recognition of mycorrhizal (Myc) factors. Chitin elicitor receptor kinase 1 of Oryza sativa subsp. Japonica (OsCERK1) plays a bifunctional role in mediating both chitin-triggered immunity and symbiotic relationships with AM fungi. However, it remains unclear whether OsCERK1 can directly recognize chitin molecules. In this study, we show that OsCERK1 binds to the chitin hexamer ((NAG)₆) and tetramer ((NAG)₄) directly and determine the crystal structure of the OsCERK1-(NAG)₆ complex at 2 Å. The structure shows that one OsCERK1 is associated with one (NAG)₆. Upon recognition, chitin hexamer binds OsCERK1 by interacting with the shallow groove on the surface of LysM2. These structural findings, complemented by mutational analyses, demonstrate that LysM2 is crucial for recognition of both (NAG)₆ and (NAG)₄. Altogether, these findings provide structural insights into the ability of OsCERK1 in chitin perception, which will lead to a better understanding of the role of OsCERK1 in mediating both immunity and symbiosis in rice.     

山楂粉蝶NPV的形态发生及其宿主细胞器的超微病理变化

应用电子显微技术,对山楂粉蝶核型多角体病毒感染三龄幼虫后,该病毒在体内的形态发生过程及其宿主细胞病理超微结构的变化等进行了观察与分析。结果表明：病毒感染７２～１６８ｈ后,山楂粉蝶幼虫中肠上皮细胞内出现病毒发生基质、核衣壳、套膜、丝状纤维或称前多角体蛋白。病毒粒子、病毒束,以及病毒束嵌入多角体蛋白,装配成完全的病毒多角体。在相应的中肠上皮细胞内,细胞器如线粒体、粗面内质网、核糖体、溶酶体等均发生显著的病理变化。同时还应指出的是内质网呈指纹图谱型,板层体与示髓样小体等膜状结构的病理变化比较特殊。     

Nonreductive release of O-linked oligosaccharides from mucin glycoproteins for structure/function assignments as neoglycolipids: application in the detection of novel ligands for E-selectin

The neoglycolipid technology comprises several microproceduresinvolving the generation of lipid-linked oligosaccharide probesfor carbohydrate recognition studies in conjunction with oligosaccharidesequence determination by mass spectrometry. Although applicableto any desired oilgosaccharides, procedures are greatly facilitatedif the ohgosaccharides are nonreduced, as conjugation is byreductive amination of a reducing end aldehyde to a phosphatidylethanolamine.Using bovine submaxillary mucin as a model for release of O-glycansin the reducing state, and based on yields of neoglycolipidsand side-products from "peeling" reactions and degradation,aqueous ethylamine 70% w/v at 22°C for 48 h has been selectedin preference to other conditions, triethylainine, sodium hydroxide,and bydrazine. The integrity of the main acidic and neutraloligosaccharides released under these conditions, di- to octasaccharides,was established by analyses of free oligosaccharides by liquidsecondary ion mass spectrometry (LSIMS) and of the derived neoglycolipidsby TLCLSIMS; the repertoire compared favorably with that ofthe oligosaccharide alditols generated by conventional reductivealkaline borohydride treatment. More forcing conditions of ethylamine70% w/v at 65°C for 6 h were required to release oligosaccharidesfrom porcine gastric mucin; di- to nonasaccharides were obtainedof which about one-third had an intact core GalNAc. Relativeto yields after reductive alkaline hydrolysis, the overall yieldsfor these two glycoproteins were 20% and 40–50% for acidicand neutral oligosaccharides, respectively. Among O-glycansreleased from an ovarian cystadenoma glycoprotein using ethylamine,three variants of the sulfated Le^a/x sequences were identifiedas ligands for the endothelial adhesion molecule E-selectin,one of which is based on the unusual backbone Gal-3/4GlcNAc-3Gal-3Gal. mucins O-linked oligosaccharides TLC-LSIMS neoglycolipids E-selectin     

Studying dynamic process of photosensitization by ESR method

By kinetic methods, functional relation of TAN radical, produced in the process of TEMPONE trapping O2, vs. time during photosensitization was established. Accordingly relative rate constants of generating all kinds of active intermediates through types I and II mechanism of photosensitization can be calculated. Using the formula and experimental results, the relative rate constants of generating O2, O2 and PS2 of three kinds of perylenequinone photosensitizer: HA, HB and CP in DMF-H20 and DMSO-H2O system were calculated, and then the structure-activity relationship of perylenequinone photosensitizer and the relation between photosensitivity and solvent was studied.     

诺卡氏菌对油酸的羟基化作用

诺卡氏菌(Nocardia sp．N13)休止细胞对油酸(顾-9-十八碳烯酸)进行羟基化作用。用休止细胞转化时最适温度为30℃,最适pH为6．5,当菌体(干菌)与底物的最适比例为20 ：1对,对油酸的转化率为83．4％,不同的表面活性剂影响转化的效果。N13休止细胞在磷酸缓冲液中重复转化3次仍可保持50％左右的转化率。红外光谱、质谱、棱磁共振鉴定产物为10－羟基硬脂酸。     

Molecular forms of acetylcholinesterase present in the white and grey matter of pig brain

Extraction of the white matter of pig brain with EDTA, lysolecithin or Triton X-100 gave poor yields of soluble acetylcholinesterase although these agents had proved effective at solubilizing the enzyme in the grey matter. This finding, together with the observation that the strong detergent sodium deoxycholate, was needed to solubilize the enzyme, shows that it is more difficult to remove acetylcholinesterase from the white matter of brain than from the grey. This could mean that the enzyme in the white matter is more firmly bound to the membrane than the enzyme in the grey matter.The difference in binding of the enzyme from the two regions of the brain is also reflected in the affinity chromatography experiments which showed a lower recovery for the acetylcholinesterase of white matter compared with the enzyme from grey matter.Starch-block electrophoresis of acetylcholinesterase showed a single negatively charged peak of activity for both the naturally soluble and the deoxycholate solubilized preparations. The presence of only one form on electrophoresis suggests that the molecular species of acetylcholinesterase do not arise from differences in charge.Sucrose density gradient centrifugation of the two preparations from white matter gave a single peak of activity with a sedimentation constant of about 10 S. This corresponds closely to the major species of molecular weight 260,000 detected by gradient gel electrophoresis. Other forms detected in both enzyme preparations by gradient gel electrophoresis were species with molecular weights of 660,000, 180,000, 130,000 and 115,000. The significance of these species in terms of the formation of oligomers is discussed.A comparison was made with the corresponding preparations of acetylcholinesterase from the grey matter and the results showed that acetylcholinesterase from the white and grey matter of pig brain were very similar. The exception to this was the species with a molecular weight of 68,000 which was present in the grey but not the white matter of pig brain.