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Proteome analysis of round-headed and normal spermatozoa by 2-D fluorescence difference gel electrophoresis and mass spectrometry 被引量:9
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作者 Ting-Ting Liao Zhen Xiang Wen-Bing Zhu Li-Qing Fan 《Asian Journal of Andrology》 SCIE CAS CSCD 2009年第6期683-694,共12页
Globozoospermia is a severe form of teratozoospermia characterized by round-headed spermatozoa with an absent acrosome, an aberrant nuclear membrane and midpiece defects. Globozoospermia is diagnosed by the presence o... Globozoospermia is a severe form of teratozoospermia characterized by round-headed spermatozoa with an absent acrosome, an aberrant nuclear membrane and midpiece defects. Globozoospermia is diagnosed by the presence of 100% round-headed spermatozoa on semen analysis, and patients with this condition are absolutely infertile. The objective of this study was to investigate the differences in protein expression between human round- headed and normal spermatozoa. Two-dimensional (2-D) fluorescence difference gel electrophoresis (DIGE) coupled with mass spectrometry (MS) was used in this study. Over 61 protein spots were analysed in each paired normal/round-headed comparison, using DIGE technology along with an internal standard. In total, 35 protein spots identified by tandem mass spectrometry (MS/MS) exhibited significant changes (paired t-test, P 〈 0.05) in the expression level between normal and round-headed spermatozoa. A total of nine proteins were found to be upregulated and 26 proteins were found to be downregulated in round-headed spermatozoa compared with normal spermatozoa. The differentially expressed proteins that we identified may have important roles in a variety of cellular processes and structures, including spermatogenesis, cell skeleton, metabolism and spermatozoa motility. 展开更多
关键词 differential protein GLOBOZOOSPERMIA mass spectrometry (MS) two-dimensional difference gel electrophoresis (2-D DIGE)
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Agronomic Trait and Protein Component of F_2 Hybrid Originated from Intergeneric Somatic Hybridization Between Triticum aestivum and Agropyron elongatum 被引量:4
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作者 向凤宁 冯保民 +1 位作者 夏光敏 陈惠民 《Acta Botanica Sinica》 CSCD 2001年第3期232-237,共6页
Protoplasts derived from common wheat (Triticum aestivum L,. cv. Jinan 177) were fused with UV-treated protoplasts of Agropyron elongatum. (Host) Nevski by PEG method, and fertile asymmetric somatic hybrid plants rese... Protoplasts derived from common wheat (Triticum aestivum L,. cv. Jinan 177) were fused with UV-treated protoplasts of Agropyron elongatum. (Host) Nevski by PEG method, and fertile asymmetric somatic hybrid plants resembling wheat morphology were obtained. The F-2 hybrid plants could be divided into 3 types according to their morphology. Type I hybrids had high and loosely standing stalks with big spikes and grains. Type ii hybrids were dwarf and compact in shape with high tillering ability and smaller spikes. Type III hybrids were similar to type I as a whole but had more compact and erect spikes. All the F-2 hybrid lines were superior to wheat in seed protein content, although some difference existed between themselves. Protein analysis of immature embryos and flag leaves from hybrids by two-dimensional electrophoresis showed that they possessed characteristic proteins of both parents and some new proteins. There existed also some different kinds of proteins in different lines. 展开更多
关键词 F-2 somatic hybrid plant Triticum aestivum Agropyron elongatum two-dimensional electrophoresis crude protein protein component
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Combining Phytate/Ca^(2+) Fractionation with Trichloroacetic Acid/Acetone Precipitation Improved Separation of Low-Abundant Proteins of Wheat (Triticum aestivum L.) Leaf for Proteomic Analysis 被引量:1
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作者 Muhammad A R F Sultan LIU Hui +2 位作者 CHENG Yu-Feng ZHANG Pei-pei ZHAO Hui-xian 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2013年第7期1123-1129,共7页
Proteomic assessment of low-abundance leaf proteins is hindered by the large quantity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) present within plant leaf tissues. In the present study, total prote... Proteomic assessment of low-abundance leaf proteins is hindered by the large quantity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) present within plant leaf tissues. In the present study, total proteins were extracted from wheat (Triticum aestivum L.) leaves by a conventional trichloroacetic acid (TCA)/acetone method and a protocol first developed in this work. Phytate/Ca2+ fractionation and TCA/acetone precipitation were combined to design an improved TCA/acetone method. The extracted proteins were analysed by two-dimensional gel electrophoresis (2-DE). The resulting 2-DE images were compared to reveal major differences. The results showed that large quantities of Rubisco were deleted from wheat leaf proteins prepared by the improved method. As many as (758±4) protein spots were detected from 2-DE images of protein extracts obtained by the improved method, 130 more than those detected by the TCA/acetone method. Further analysis indicated that more protein spots could be detected at regions of pI 4.00-4.99 and 6.50-7.00 in the improved method-based 2-DE images. Our findings indicated that the improved method is an efficient protein preparation protocol for separating low-abundance proteins in wheat leaf tissues by 2-DE analysis. The proposed protocol is simple, fast, inexpensive and also applicable to protein preparations of other plants. 展开更多
关键词 Triticum aestivum L. RUBISCO low-abundance protein phytate/Ca2 two-dimensional gel electrophoresis plant leaf proteomics
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CHANGES OF NUCLEAR MATRIX PROTEIN AND ITS RELATIONSHIP WITH c-erbB-2 IN HUMAN COLON ADENOCARCINOMA
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作者 王娅兰 高静 李圆圆 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2005年第4期255-259,共5页
Objective: Nuclear matrix protein is tissue, cell-type specific, and tumor-relative. It plays an important role in the regulation of intranuclear processes. Some researches also showed that a c-erbB-2 promoter-specif... Objective: Nuclear matrix protein is tissue, cell-type specific, and tumor-relative. It plays an important role in the regulation of intranuclear processes. Some researches also showed that a c-erbB-2 promoter-specific DNA-binding nuclear matrix protein is present only in malignant human breast tissues and induces mitogenesis and cell surface expression of the c-erbB-2 protein in resting NIH/3T3 cells. But it is not clear that how it in colon adenocarcinomas. Methods: Two-dimensional gel electrophoretic method was used for NMP identification and immunohistochemistry was used for c-erbB-2 detection in 12 cases of colon adenocarcinomas and matched adjacent normal colon tissues. Results: 5 different nuclear matrix proteins (named C1-C5) were identified in 12 colon adenocarcinoma specimens, but not in the matched adjacent normal colon tissues; 3 nuclear matrix proteins (named N1-N3) were identified in all 12 matched adjacent normal colon tissues, but not in colon adenocarcinoma specimens. A nuclear matrix protein (named N4) was detected in all of 9 moderated-well differentiated adenocarcinomas and all 12 matched adjacent normal colon tissues, but not in 3 poor-differentiated adenocarcinomas. All of the 10 colon adenocarcinomas which had the nuclear matrix protein C4 were c-erbB-2 expression positive. Conclusion: The data suggest that there are specific nuclear matrix proteins in colon adenocarcinomas and its subtypes, which maybe valuable to serve as markers of colon adenocarcinomas in future. Nuclear matrix protein C4 probably is a c-erbB-2 promotor-specific nuclear matrix protein in colon adenocarcinomas, and may induce the expression of c-erbB-2. 展开更多
关键词 Colon adenocarcinoma Nuclear matrix protein C-ERBB-2 two-dimensional gel electrophoresis
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Plant Proteomics in the Post-genomic Era 被引量:12
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作者 郭奕明 沈世华 +1 位作者 荆玉祥 匡廷云 《Acta Botanica Sinica》 CSCD 2002年第6期631-641,共11页
Proteomics is one of the most active research fields in the post-genomic era. Here we briefly introduce the scientific background of the origination of proteomics and its content, research method. The new developments... Proteomics is one of the most active research fields in the post-genomic era. Here we briefly introduce the scientific background of the origination of proteomics and its content, research method. The new developments of proteomics at the levels of individual plants, tissues, organs and organells, as well as its applications in the area of plant genetic diversity, mutant characterization, and plant physiology, etc are reviewed. At last, the challenge and prospect of proteomics are discussed. 展开更多
关键词 genome post-genome PROTEOME plant proteomics matrix-assisted laser-desorption/ionizationtime-of-flight mass spectrometry (MALDI-TOF/MS) two-dimensional polyacrylamide gel electrophoresis (2D-PAGE)
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Proteomic Analysis of Wheat Seed in Response to Drought Stress 被引量:2
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作者 ZHANG Yu-feng HUANG Xiu-wen +4 位作者 WANG Li-li WEI Liu WU Zhi-hui YOU Ming-shan LI Bao-yun 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2014年第5期919-925,共7页
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... 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. 展开更多
关键词 wheat seed drought stress two-dimensional electrophoresis (2-de)
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Identification of differentially expressed proteins in poplar leaves in-duced by Marssonina brunnea f. sp. Multigermtubi 被引量:6
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作者 Kun Yuan Bo Zhang +3 位作者 Yanmei Zhang Qiang Cheng Mingxiu Wang Minren Huang 《Journal of Genetics and Genomics》 SCIE CAS CSCD 北大核心 2008年第1期49-60,共12页
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) in... 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. 展开更多
关键词 black spot disease defense response two-dimensional gel electrophoresis (2-de) matrix assisted laser desorption/ionization time of flightmass spectrometry (MALDI-TOF MS) POPLAR
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5-Aminolevulinic acid alleviates herbicide-induced physiological and ultrastructural changes in Brassica napus 被引量:2
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作者 XU Ling Faisal Islam +2 位作者 ZHANG Wen-fang Muhammad A Ghani Basharat Ali 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2018年第3期579-592,共14页
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 ultrastruct... 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. 展开更多
关键词 Brassica napus L. ALA ZJ0273 plant growth antioxidant enzyme activities ULTRASTRUCTURE two-dimensional gel electrophoresis (2-de)
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Identification of the major allergen of Macrobrachium rosenbergii (giant freshwater prawn) 被引量:1
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作者 Zailatul Hani Mohamad Yadzir Rosmilah Misnan +3 位作者 Noormalin Abdullah Faizal Bakhtiar Masita Arip Shahnaz Murad 《Asian Pacific Journal of Tropical Biomedicine》 SCIE CAS 2012年第1期50-54,共5页
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... 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. 展开更多
关键词 MACROBRACHIUM rosenbergii Major ALLERGEN MALDI-TOF TROPOMYOSIN ARGININE kinase SDS-PAGE Immunoblotting 2-de electrophoresis IgE reactivity
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Differential Proteomic Analysis of Neonatal Cardiomyocytes in Response to β-Adrenergic Receptor Stimulation
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作者 LI Zi-jian LIU Ning +3 位作者 LIU Zhi-qiang LIU Shu-ying HAN Qi-de ZHANG You-yi 《Chemical Research in Chinese Universities》 SCIE CAS CSCD 2006年第3期319-323,共5页
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, proteom... 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. 展开更多
关键词 PROTEOME two-dimensional electrophoresis(2-de) MALDI-MS Β-ADRENOCEPTOR CARDIOMYOCYTE
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Application of proteome technology in screening biomarkers associated with gastric cancer
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作者 Chibo Liu Yong Liang Haibao Wang Chunqin Pan 《The Chinese-German Journal of Clinical Oncology》 CAS 2008年第7期394-397,共4页
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 ... 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. 展开更多
关键词 PROTEOME two-dimensional gel electrophoresis (2-de) gastric carcinoma tumor markers BIOLOGY
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Comparative analysis of bed density, total phenol content and protein expression pattern in <i>Posidonia oceanica</i>(L.) Delile
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作者 Alice Rotini Letizia Anello +3 位作者 Maria Di Bernardo Agata Giallongo Luigi Valiante Luciana Migliore 《Open Journal of Ecology》 2013年第6期438-444,共7页
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 ... 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. 展开更多
关键词 POSIDONIA oceanica Rhizome Seagrass Monitoring Total Phenols 2-D electrophoresis (2-de) BED DENSITY PROTEIN Expression Pattern
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Effect of high salinity on cell growth and protein production of Antarctic ice microalgae Chlamydomonas sp. ICE-L
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作者 郑洲 缪锦来 +5 位作者 阚光峰 金青 丁燏 刘芳明 王守强 王以斌 《Chinese Journal of Polar Science》 2010年第1期81-90,共10页
Antarctic ice microalgae Chlamydomonas sp.ICE-L can survive and thrive in Antarctic sea ice.In this study,Chlamydomonas sp.ICE-L could survive at the salinity of 132‰ NaCl.SDS-PAGE showed that the density of 2 bands... Antarctic ice microalgae Chlamydomonas sp.ICE-L can survive and thrive in Antarctic sea ice.In this study,Chlamydomonas sp.ICE-L could survive at the salinity of 132‰ NaCl.SDS-PAGE showed that the density of 2 bands(26 and 36 kD) decreased obviously at the salinity of 99‰ NaCl compared to at the salinity of 33‰ NaCl.The soluble proteins in Chlamydomonas sp.ICE-L grown under salinity of 33‰ and 99% NaCl were compared by 2-D gel electrophoresis.After shocking with high salinity,8 protein spots were found to disappear,and the density of 28 protein spots decreased.In addition,19 protein spots were enhanced or induced,including one new peptide(51 kD).The changes of proteins might be correlated with the resistance for Chlamydomonas sp.ICE-L to high salinity. 展开更多
关键词 Antarctic ice alga Chlamydomonas sp.ICE-L soluble proteins two-dimensional polyacrylamide gel electrophoresis gel electrophoresis(2-D PAGE) analysis high-salinity press.
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Increased RhoGDI2 and peroxiredoxin 5 levels in asthmatic murine model of β2-adrenoceptor desensitization: A proteomics approach 被引量:14
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作者 LIU Hua ZHOU Lin-fu ZHANG Qian QIAN Fen-hong YIN Kai-sheng HUANG Mao ZHANG Xi-long 《Chinese Medical Journal》 SCIE CAS CSCD 2008年第4期355-362,共8页
Background β2-adrenoceptor (β2AR) desensitization is a common problem in clinical practice, β2AR desensitization proceeds by at least such three mechanisms as heterologous desensitization, homologous desensitizat... Background β2-adrenoceptor (β2AR) desensitization is a common problem in clinical practice, β2AR desensitization proceeds by at least such three mechanisms as heterologous desensitization, homologous desensitization and a kind of agonist-induced rapid phosphorylation by a variety of serine/threonine kinases. It is not clear whether there are other mechanisms, This study aimed to investigate potential mechanisms of β2AR desensitization.Methods Twenty-four BALB/c (6-8 weeks old) mice were divided into three groups, which is, group A, phosphate buffered saline (PBS)-treated; group B, ovalbumin (OVA)-induced; and group C, salbutamol-treated. Inflammatory cell counts, cytokine concentrations of bronchoalveolar lavage fluid (BALF), pathological sections, total serum IgE, airway responsiveness, membrane receptor numbers and total amount of β2AR were observed. Asthmatic mouse model and β2AR desensitization asthmatic mouse model were established. Groups B and C were selected for two-dimensional gel electrophoresis (2DE) analysis so as to find key protein spots related to β2AR desensitization.Results Asthmatic mouse model and β2AR desensitization asthmatic mouse model were verified by inflammatory cell count, cytokine concentration of BALF, serum IgE level, airway hyperreactivity measurement, radioligand receptor binding assay, Western blot analysis, and pathologic examination. Then the two groups (groups B and C) were subjected to 2DE. Two key protein spots associated with β2AR desensitization, Rho GDP-dissociation inhibitor 2 (RhoGDl2) and peroxiredoxin 5, were found by comparative proteomics (2DE and mass spectrum analysis).Conclusion Oxidative stress and small G protein regulators may play an important role in the process of β2AR desensitization.v 展开更多
关键词 β2-adrenoceptor desensitization two-dimensional gel electrophoresis mass spectrometry peroxiredoxin 5 RhoGDP-dissociation inhibitor 2
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Science Letters:Proteomic analysis of differentially expressed proteins in mice with concanavalin A-induced hepatitis 被引量:1
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作者 Xu-fei TAN Feng CHEN Shan-shan WU Yu SHI Dong-cheng LIU Zhi CHEN 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2010年第3期221-226,共6页
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. ... 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. 展开更多
关键词 Concanavalin A (Con A) Liver injury Protein two-dimensional electrophoresis (2-de)
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The Proteomic Research of the Cure of Experimental Diabetes Deafness by Granules of Eliminating Phlegm and Removing Blood Stasis 被引量:3
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作者 郭宏 熊大经 《Journal of Traditional Chinese Medicine》 SCIE CAS CSCD 2011年第2期88-97,共10页
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 iden... 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. 展开更多
关键词 Granules of Eliminating Phlegm and Removing Blood Stasis also known as GEPRB Diabetes deafness also known as DD two-dimentional electrophoresis also known as 2-de Matrix-assisted laser desorption ionization-time of flight- mass Spectrometry also known as MS Serum proteomics
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Proteomics analysis of cerebral cortex in Wistar rats
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作者 Xiaofeng ZHAO Jingrong WEN +1 位作者 Shu WANG Xuemin SHI 《Frontiers in Biology》 CSCD 2008年第4期419-427,共9页
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 di... 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. 展开更多
关键词 brain proteomics two-dimensional gel electrophoresis(2-de) matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS)
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