Identification of differentially expressed proteins in poplar leaves in-duced by Marssonina brunnea f. sp. Multigermtubi

Black spot disease in poplar is a disease of the leaf caused by fungus. The major pathogen is Marssonina brunnea f. sp. multigermtubi. To date, little is known about the molecular mechanism of poplar （M. brunnea） interaction. In order to identify the proteins related to disease resistance and understand its molecular basis, the clone ＂NL895＂ （P. euramericana CL＂NL895＂）, which is highly resistant to M. brunnea f. sp. multigermtubi, was used in this study. We used two-dimensional gel electrophoresis （2-DE） and mass spectrometry （MS） to identify the proteins in poplar leaves that were differentially expressed in response to black spot disease pathogen, M. brunnea f. sp. multigermtubi. Proteins extracted from poplar leaves at 0, 12, 24, 48, and 72 h after pathogen-inoculation were separated by 2-DE, About 500 reproducible protein spots were detected, of which 40 protein spots displayed differential expression in levels and were subjected to Matrix assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF MS） followed by database searching. According to the function, the identified proteins were sorted into five categories, that is, protein synthesis, metabolism, defense response and unclassified proteins.

5-Aminolevulinic acid alleviates herbicide-induced physiological and ultrastructural changes in Brassica napus

It is well known that application of 5-aminolevuJinic acid （ALA） could promote the plant growth under abiotic stress in oilseed rape （Brassica napus L.）. However, the specifics of its physiological and ultrastructural regulation under herbi- cide stress conditions are poorly understood. In the present study, alleviating role of ALA in B. napus was investigated under four levels of herbicide propyl 4-（2-（4,6-dimethoxypyrimidin-2-yloxy） benzylamino） benzoate （ZJ0273） （0, 100, 200 and 500 mg L-1） with or without 1 mg L-1 ALA treated for 48 or 72 h. Results showed that after 48 h of herbicide stress, the growth of rape seedlings was significantly inhibited with the successive increases of the ZJ0273 concentrations from 0 to 500 mg L-1, but this inhibition was obviously alleviated by exogenous application of ALA. However, when treatment time prolonged to 72 h, the recovery effects of ALA could not be evaluated due to the death of plants treated with the highest concentration of ZJ0273 （500 mg L-1）. Further, the root oxidizability and activities of antioxidant enzymes （superoxide dismutase, perox- idase and ascorbate peroxidase） were dramatically enhanced by the application of 1 mg L-1 ALA under herbicide stress. Therefore, plants treated with ALA dynamically modulated their antioxidant defenses to reduce reactive oxygen species （ROS） accumulation and malondialdehyde （MDA） content induced by herbicide stress. Additionally, exogenously applied ALA improved the ultrastructure＇s of chloroplast, mitochondria and nucleus, and induced the production of stress proteins. Our results suggest that ALA could be considered as a potential plant growth regulator for the improvement of herbicide tolerance through alleviation of the physiological and ultrastructural changes induced by the herbicide in crop production.

Proteomic Analysis of Wheat Seed in Response to Drought Stress

Drought stress is one of the major factors affecting in wheat yield and grain quality. In order to investigate how drought stress might influence wheat quality during grain filling, three wheat cultivars Gaocheng 8901, Jagger and Nongda 3406 were subjected to drought stress during the grain filling stage. Neither globulin and glutenin, nor the relative percentage ofamylose significantly changed following drought treatments, whereas albumin and gliadin concentrations did. The SDS-sedimentation, which has a strong linear correlation with wheat baking quality was markedly decreased following drought stress. These results indicated that drought had an adverse effect on wheat qualit3,. In order to investigate the protein complexes in the wheat flour, the data from native PAGE and SDS-PAGE were combined and a total of 14 spots were successfully identified, and of these eight protein types were determined to be potential complex forming proteins.

Identification of the major allergen of Macrobrachium rosenbergii (giant freshwater prawn)

Objective:To characterize the major allergens of Macrobrachium rosenbergii.(giant freshwater prawn).Methods:Raw and cooked extracts of the giant freshwater prawn were prepared.The IgE reactivity pattern was identified by sodium dodecyl sulfate polyaerylamide gel electrophoresis(SDS-PAGE)and immunoblotting technique with the sera of 20 skin prick test(SPT)positive patients.The major allergen identified was then characterized using the proteomics approach involving a combination of two-dimensional(2-DE)electrophoresis,mass spectrometry and bioinformatics tools.Results:SDS-PAGE of the raw extract showed 23 protein bands(15-250 kDa)but those ranging from 40 to 100 kDa were not found in the cooked extract.From immunoblotting experiments,raw and cooked extracts demonstrated 11 and 5 IgE-binding proteins,respectively,with a molecular mass ranging from 15 to 155 kDa.A heat-resistant 36 kDa protein was identified as the major allergen of both extracts.In addition,a 42 kDa heat-sensitive protein was shown to be a major allergen of the raw extract.The 2-DE gel fractionated the prawn proteins to more than 50 different protein spots.Of these,10 spots showed specific:IgE reactivity with patients'sera.Matrix assisted laser desorption/ionization-lime of flight(MALDI-TOF)analysis led to identification of 2 important allergens,tropomyosin and arginine kinase.Conclusions:It can be concluded that the availability of such allergens would help in component-based diagnosis and therapy of prawn allergies.

Differential Proteomic Analysis of Neonatal Cardiomyocytes in Response to β-Adrenergic Receptor Stimulation

β-Adrenoceptors（β-ARs） play a critical role in regulating cardiac functions under both physiological and pathological conditions. To further explore the mechanisms through which β-ARs perform its actions, proteomic approaches were adopted to study the global protein patterns in cultured neonatal rat cardiomyocytes exposed to iseproterenol （ISO）. A modified method, ＂Mirror Images in One Gel＂, was used to improve the reproducibility and resolution power of two-dimensional electrophoresis. A 2-DE map with a good reproducibility was obtained in which 1281 ± 70 spots were detected and about 1191± 54 spots were matched, with an average matching rate of 92. 9%. Nine proteins with significant changes were identified by using peptide mass fingerprinting（PMF） data obtained vht MALDI-MS.

Application of proteome technology in screening biomarkers associated with gastric cancer

Objective: To initially explore the application of proteome technologies in study of serum, to establish two-dimen-sional gel electrophoresis (2-DE) profiles of human gastric cancer serum and paired normal serum, and to screen and identify differentially expressed proteins in poorly-differentiated gastric cancer serum and paired normal serum, in which try to find out significant biomarker candidates for gastric cancer. Methods: 2-DE was adopted to separate the total proteins of poorly-differentiated gastric cancer serum and paired normal serum. After staining and analyzing by ImageMaster 2D Elite software, the differentially expressed proteins were identified by matrix-assisted laser desorption/ionization-time of flight- mass spectrometry (MALDI-TOF-MS). Results: 2-DE serum profiles with high resolution were obtained. Five protein spots were found as differentially-expressed proteins and identified as Serpin B6 (Placental thrombin inhibitor) Cytoplasmic antiproteinase (CAP) (Protease inhibitor6) (PI-6), Septin-1 (LARP) (Serologically defined breast cancer antigen NY-BR-24), Kallikrein-6 precursor (Protease M) (Neurosin) (Zyme) (SP59), Hemoglobin beta chain, Hemoglobin beta chain and Beta-defensin 108 precursor (Beta-defensin 8) (DEFB-8). Conclusion: The differential proteins were demonstrated to present in poorly-differentiated gastric cancer serum and paired normal serum. The molecular biomarkers associated with poorly differentiated gastric cancer could be possibly identified by the high throughput screening proteome technology.

Comparative analysis of bed density, total phenol content and protein expression pattern in <i>Posidonia oceanica</i>(L.) Delile

Posidonia oceanicameadows are experiencing a progressive decline, and monitoring their status is crucial for the maintenance of theseecosystems. We performed a comparativeanalysis of bed density, total phenol content and protein expression pattern to assess the conservation status ofPosidoniaplants from the S. Marinella (Rome, Italy) meadow. The total phenol content was inversely related to maximum beddensity, confirming the relationship betweenhigh phenol content and stressful conditions. In addition, protein expression pattern profilesshowed that the number of differentially expressed proteins was dramatically reduced in the latest years compared to previous analyses. Our results support the usefulness of integrating solid descriptors, such as phenol content, with novel biochemical/molecular approaches in the monitoring of meadows.

Science Letters:Proteomic analysis of differentially expressed proteins in mice with concanavalin A-induced hepatitis

Objective： To find new protein biomarkers for the detection and evaluation of liver injury and to analyze the relationship between such proteins and disease progression in concanavalin A （Con A）-induced hepatitis. Methods： Twenty-five mice were randomly divided into five groups： an untreated group, a control group injected with phosphate buffered saline （PBS）, and groups with Con A-induced hepatitis evaluated at 1, 3 and 6 h. Two-dimensional gel electrophoresis （2-DE） and mass spectrometry （MS） were used to identify differences in protein expression among groups. Quantitative real-time polymerase chain reaction （qRT-PCR） was performed to verify the results. Results： In mice with Con A-induced hepatitis, expression levels of four proteins were increased： RIKEN, fructose bisphosphatase 1 （fbp1）, ketohexokinase （khk）, and Chain A of class pi glutathione S-transferase. Changes in fbpl and khk were confirmed by qRT-PCR. Conclusion： Levels of two proteins, fbp1 and khk, are clearly up-regulated in mice with Con A-induced hepatitis.

The Proteomic Research of the Cure of Experimental Diabetes Deafness by Granules of Eliminating Phlegm and Removing Blood Stasis

Objective: Observing the expression changes of serum proteome in model rats after intervention of the Granules of Eliminating Phlegm and Removing Blood Stasis (豁痰祛瘀颗粒 also known as GEPRB), screening out and identifying the differentially expressed proteins by mass spectrometry and bioinformatics analysis, discussing the molecular mechanism of control the Diabetes deafness by GEPRB. Methods: By use of proteomics technology, the serum protein serum proteome of the control group, model control group, Duxil and each observation group were observed for 2-DE gel pattern matching, and the difference in the relative content of 2 times was chosen for the differentially expressed proteins. Identification of differentially expressed proteins by MALDI-TOF MS/MS, the authors further analysis the phosphorylation, subcellular localization, interaction, direct regulation, and transmembrane of the differences proteins by the way of bioinformatics analysis. Sixty SPF level SD rats elected in diabetic rats model group (abbreviated as DM group) were be randomly divided into 5 groups based on random number sheet, namely model control group, positive drug control group (Du-ke-xi group) and Mai-tong-fang high, medium and low dose group respectively. In addition, set of normal control group. 10 rats in each group. Results: By Coomassie brilliant blue staining, identified 51 differential protein spots dug from 2-D gel by mass spectrometry, successfully identified 13 non-redundant proteins. Most of the identified proteins were secreted protein and belong to different protein families. There were about 12 proteins have the transmembrane region from the authors’ result, ten of them were plasma membrane proteins. Conclusion: It’s suggesting that 13 differential proteins is most likely the protein response to GEPRB in vivo, these proteins may play key role for the treatment of GEPRB to Diabetes deafness. The two highly differentially expressed proteins Apolipoprotein E (apoE) and C3 may be a potential drug target of GEPRB.

Proteomics analysis of cerebral cortex in Wistar rats

To analyze the protein expression pattern of the cerebral cortex in Wistar rats using the proteomics approach,proteins were separated by two-dimensional gel electrophoresis,stained with Coomassie brilliant blue and digested with trypsin.Then,we analyzed the peptide section using a matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS)and identified the protein by indexing special database(SwissProt)according to the finger printing of the peptide quality.Eighty-four protein spots were identified,including metabolic enzymes,skeleton proteins,heat shock proteins,antioxidant proteins,signaling proteins,proteasome related proteins,neuron and glial specific proteins and serum associated proteins.The result of this study enriches the database of the proteome in the cerebral cortex of rats and lays a foundation for further research of neurological disorders in rat models.