Post-translational tyrosine nitration of eosinophil granule toxins mediated by eosinophil peroxidase

Nitration of tyrosine residues has been observed during various acute and chronic inflammatory diseases. However, the mechanism of tyrosine nitration and the nature of the proteins that become tyrosine nitrated during inflammation remain unclear. Here we show that eosinophils but not other cell types including neutrophils contain nitrotyrosine-positive proteins in specific granules. Furthermore, we demonstrate that the human eosinophil toxins, eosinophil peroxidase (EPO), major basic protein, eosinophil-derived neurotoxin (EDN) and eosinophil cationic protein (ECP), and the respective murine toxins, are post-translationally modified by nitration at tyrosine residues during cell maturation. High resolution affinity-mass spectrometry identified specific single nitration sites at Tyr349 in EPO and Tyr33 in both ECP and EDN. ECP and EDN crystal structures revealed and EPO structure modeling suggested that the nitrated tyrosine residues in the toxins are surface exposed. Studies in EPO(-/-), gp91phox(-/-), and NOS(-/-) mice revealed that tyrosine nitration of these toxins is mediated by EPO in the presence of hydrogen peroxide and minute amounts of NOx. Tyrosine nitration of eosinophil granule toxins occurs during maturation of eosinophils, independent of inflammation. These results provide evidence that post-translational tyrosine nitration is unique to eosinophils.     

Synthesis of rhamnogalacturonan I fragments by a modular design principle

The improved syntheses of methyl 2-O-acetyl-3-O-benzyl-alpha-L-rhamnopyranoside (12) and 1,2-di-O-acetyl-3-O-benzyl-alpha-L-rhamnopyranose (15), which were used as glycosyl acceptor and donor, respectively, are described. Glycosylation of the O-4 position of both rhamnose derivatives with 2,3,4,6-tetra-O-benzoyl-alpha-D-galactopyranosyl bromide (26) provided disaccharides 27 and 29. After partial deprotection of 27 and coupling of the resulting 28 with disaccharide 19, tetrasaccharide 31 was obtained. Furthermore, transforming of 29 into the corresponding bromide 30 and coupling with galacturonates 16 and 32 provided trisaccharides 33 and 34, respectively, which could be regarded as building blocks of ramified rhamnogalacturonan fragments. The preparation of tetra- (21) and hexasaccharide (25) of rhamnogalacturonan I is reported to demonstrate the feasibility of the synthesis of larger pectin fragments using the modular design principle with this type of building blocks.     

The AAE14 gene encodes the Arabidopsis o-succinylbenzoyl-CoA ligase that is essential for phylloquinone synthesis and photosystem-I function

Phylloquinone is the one-electron carrier at the A₁ site of photosystem I, and is essential for photosynthesis. Arabidopsis mutants deficient in early steps of phylloquinone synthesis do not become autotrophic and are seedling lethals, even when grown on sucrose-supplemented media. Here, we identify acyl-activating enzyme 14 (AAE14, At1g30520) as the o -succinylbenzoyl-coenzyme A (OSB-CoA) ligase acting in phylloquinone synthesis. Three aae14 mutant alleles, identified by reverse genetics, were found to be seedling lethal, to contain no detectable phylloquinone (< 0.1 pmol mg⁻¹ fresh weight) compared with 10 pmol mg⁻¹ fresh weight in wild-type leaves, and to accumulate OSB. AAE14 was able to restore menaquinone biosynthesis when expressed in an Escherichia coli mutant disrupted in the menE gene that encodes the bacterial OSB-CoA ligase. Weak expression of an AAE14 transgene in mutant plants (controlled by the uninduced XVE promoter) resulted in chlorotic, slow-growing plants that accumulated an average of 4.7 pmol mg⁻¹ fresh weight of phylloquinone. Inducing the XVE promoter in these plants, or expressing an AAE14 transgene under the control of the CaMV 35S promoter, led to full complementation of the mutant phenotype. aae14 -mutant plants were also able to synthesize phylloquinone when provided with 1,4-dihydroxy-2-naphthoate, an intermediate in phylloquinone synthesis downstream of the OSB-CoA ligase reaction. Expression of an AAE14:GFP reporter construct indicated that the protein accumulated in discrete foci within the chloroplasts. This and other evidence suggests that the enzymes of phylloquinone synthesis from isochorismate may form a complex in the chloroplast stroma to facilitate the efficient channeling of intermediates through the pathway.     

Ectopic Expression of the Pttknl Gene Induces Alterations in the Morphology of the Leaves and Flowers in Petunia hybrida Vilm.

A novel knottedl-like homeobox （knox） gene, Pttknl （Populus tremulaxtremuloides knottedl）, isolated from the cambial region of hybrid aspen, was introduced into Petunia hybrida Vilm. using the leafdisc method mediated by Agrobacterium. A series of novel phenotypes was observed in transgenic petunia plants, including the formation of ectopic spikes on the adaxial surface of corollas and small petals on the abaxial surface of corollas, fusion of floral organs, shortening of corolla midribs, the formation of tumor-like knots along the midrib on the abaxial surface and serrated lobs of corolla margins, and alterations in petal color; except for changes in the leaves and plant architecture, RT-PCR showed that the Pttknl gene was expressed in the leaves of different petunia transgenic plants, whereas no signal was detected in wild-type plants. The possible function of Pttknl in leaf and flower development is discussed.     

An expression profile of human α-lactalbumin in the milk of transgenic mouse

a-lactalbumin(a-Lac),amajorwheyprotein,isacalciummetalloprotein,thathasbeenfoundinallmilksstudiedsofar.ItinteractswithUDP-galactosyl-transferasetoformthelactosesynthetaseandthusmightbeakeyproteinforlactogenesis.Lactosesyn-thetaseispostulatedtobetherate-limitingenzymeforlactosebiosynthesis.Theincreaseda-Lacactivitycanproducesufficientlactosesynthetaseforthesynthesisoflactose,andinmilkyieldbydrawingwaterintomilk,sincelactoseisanosmoreactivemolecule.Transgenicswineoverexpressingbovinea-lactalbu…     

Screening and identification of Shigella flexneri 2a virulence-related genes induced after invasion of epithelial cells

~~Screening and identification of Shigella flexneri 2a virulence-related genes induced after invasion of epithelial cells1. Jin, Q., Yuan, Z., Xu, J., Wang, Y., Shen, Y., Lu, W., Wang, J., Liu, H., Yang, J., Yang, P., Zhang, X., Zhang, J., Yang, G, Wu, H., Qu, D., Dong, J., Sun, L., Xue, Y, Zhao, A., Gao, Y., Zhu, J., Kan, B., Ding, K.. Chen, S., Cheng, H., Yao, Z., He, B., Chen, R., Ma, D., Qiang, B., Wen, Y, Hou, Y., Yu, J., Genome sequence of Shigella flexneri 2…     

Solution structure of C-terminal Escherichia coli translation initiation factor IF2 by small-angle X-ray scattering

Initiation of protein synthesis in bacteria involves the combined action of three translation initiation factors, including translation initiation factor IF2. Structural knowledge of this bacterial protein is scarce. A fragment consisting of the four C-terminal domains of IF2 from Escherichia coli was expressed, purified, and characterized by small-angle X-ray scattering (SAXS), and from the SAXS data, a radius of gyration of 43 +/- 1 A and a maximum dimension of approximately 145 A were obtained for the molecule. Furthermore, the SAXS data revealed that E. coli IF2 in solution adopts a structure that is significantly different from the crystal structure of orthologous aIF5B from Methanobacterium thermoautotrophicum. This crystal structure constitutes the only atomic resolution structural knowledge of the full-length factor. Computer programs were applied to the SAXS data to provide an initial structural model for IF2 in solution. The low-resolution nature of SAXS prevents the elucidation of a complete and detailed structure, but the resulting model for C-terminal E. coli IF2 indicates important structural differences between the aIF5B crystal structure and IF2 in solution. The chalice-like structure with a highly exposed alpha-helical stretch observed for the aIF5B crystal structure was not found in the structural model of IF2 in solution, in which domain VI-2 is moved closer to the rest of the protein.     

早籼稻碾磨品质品种、地点、品种×地点互作效应的研究

以浙江省1995年早籼稻品种区域试验6个试点参试品种碾磨品质测定结果为试验材料,用多元分析法探讨了糙米率、精米率、整精米率3个碾磨品质的品种(基因型)、地点、品种×地点互作效应和各效应内碾磨品质间的相关关系,进行了参试品种互作效应检验。分析结果表明,糙米率、精米率以品种效应为主,整精米率性状以环境效应最大。相关分析显示,糙米率、精米率呈较强正相关,糙米率、精米率与整精米率均无相关关系。参试品种互作效应检验显示,多数品种整精米率均存在基因型×地点交互作用,糙米率、精米率多数品种较稳定。 Abstract:The genotype,test site,genotype×site effects on brown rice rate,milled rice recovery and head rice recovery as well as the correlation coefficients between the milling qualities within various effects were approached by using multiple analysis method with the data collected from early season indica rice varieties tested in regional trial in Zhejiang Province.Results showed that brown rice rate,milled rice recovery were predominantly affected by genotype.While head rice recovery was mostly conditioned by environment effect.Correlation analysis displayed that there were positive correlation between brown rice rate and milled rice rate.However,no correlation between brown rice rate,milled rice recovery and head rice recovery was found.The analysis of genotype×site of the varieties tested displayed that head rice recovery of the most tested varieties is involved with the interaction effects between the two components,while brown rice rate,milled rice recovery of the most tested varieties were more stable.     

云南地方猪种血液蛋白多态性研究

采用蛋白电泳技术研究了云南地方猪种血液蛋白多态性。共分析了3个云南地方猪种32-36个遗传位点,其中AKP、CAT、DIA、ES、G6PD、PA、6PGD、PHI、TF等9个 位点检测到多态性,多态位点百分比为0.1875-0.2121,平均杂合度为0.0712-0.1027。结果表明,云南地方猪种血液蛋白多态程度较高,反映在蛋白水平上的遗传多样性较为丰富。 Abstract:In this paper,protein electrophoresis was used to analyze the blood protein polymorphism in Yunnan local pig breeds and 32~36 genetic loci in the Yunnan local pig breeds were surveyed,Nine of them,such as AKP、CAT、DIA、ES、G6PD、PA、6PGD、PHI and TF,were found to be polymorphic,the mean heterozygosit(H)was 0.0712~0.1027.The results indicated that the blood protein polymorphism in the Yunnan local pig breeds is high,the Yunnan local pig breeds are wealthy in genetic diversity in point of their protein level.     

大口鲇和鲇鱼血清蛋白质及同工酶的比较研究

采用聚丙烯酰胺梯度凝胶垂直板电泳,分析了大口鲇和鲇鱼的血清蛋白质以及心脏、肝脏、眼和肌肉4种组织的EST及MDH同工酶。结果表明,大口鲇和鲇鱼的血清蛋白质均能分离出20条左右的谱带,两者既表现出相同的谱带,又表现出迁移率和含量都不同的带型。两者的EST和MDH同工酶在4种组织及血清中均能特异性地表达,存在明显的组织和物种特异性。本文认为肝脏是研究大口鲇和鲇鱼种群生化遗传结构与变异的理想材料,同时还探讨了两种鲇鱼的M DH同工酶位点。 Abstract:The serum proteins and isozymes in four tissues (heart,liver,eye and musele)of Smeridionalis Chen and S.asotus Linnaeus were analyzed by polyacrylamide gradient gel vertical electrophoresis.The isozymes are esterase(EST)and malate dehytrogenase(MDH).The results showed that electrophoretograme of serum proteins were about 20 protein pattens in two species catfish,they were either the same protein pattens or the different pattens.Electrophoretogram of isozymes(EST,MDH)in two species catfish indicated tissues and species specificity.Experiment considered that the liver was a good material studied biochemical genetic constitution and variation in species group of S.meridionalis Chen and S.asotus Linnaeus.