Objective To investigate the protein expression profiles of the major food‐borne pathogen Campylobacter jejuni NCTC11168.Methods Membrane and soluble cellular proteins were extracted from the genome‐sequenced C.jeju...Objective To investigate the protein expression profiles of the major food‐borne pathogen Campylobacter jejuni NCTC11168.Methods Membrane and soluble cellular proteins were extracted from the genome‐sequenced C.jejuni strain NCTC11168.Protein expression profiles were determined using two‐dimensional gel electrophoresis(2‐DE).All the detected spots on the 2‐DE map were subjected to matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry(MALDI‐TOF/TOF) analysis.Results A total of 537 and 333 spots were detected from the whole cell and membrane‐associated proteins of C.jejuni NCTC11168 cultured on Columbia agar medium at 42 ℃ by 2‐DE and Coomassie Brilliant Blue staining,respectively.Analyses of whole cell and membrane‐associated proteins included 399 and 133 spots,respectively,which included 182 and 53 functional proteins identified by MALDI‐TOF/TOF analysis.Conclusion The comprehensive expression protein profiles of C.jeuni NCTC11168 obtained in this study will be useful for elucidating the roles of these proteins in further pathogenesis investigation.展开更多
[Objective] This study aimed to explore the proteins related to pistillate flower development in different mulberry cultivars. [Method] The total proteins of the pistillate flowers of two mulberry cultivars Dal0 (Mor...[Objective] This study aimed to explore the proteins related to pistillate flower development in different mulberry cultivars. [Method] The total proteins of the pistillate flowers of two mulberry cultivars Dal0 (Morus atropurpurea Roxb.) and SG01 (Morus muIticaulis Perr.) were extracted, separated and detected through two- dimensional electrophoresis (2-DE) and mass spectrometry. [Result] There was sig- nificant difference in the expression of proteins from the pistillate flowers of different mulberry cultivars. From the 2-DE images of Dal0 and SG01, 445_+17 and 425_+12 protein spots were respectively detected. The expression levels of 75 protein spots differed significantly. Thirteen spots those were expressed at high levels and well separated were analyzed by mass spectrometry, and nine of them were identified successfully. The nine proteins are involved in the glycometabolism, protein and amino acid metabolism and defense responses during the development of mulberry pistillate flower after they were pollinated. [Conclusion] The findings will provide reference for further study on the molecular mechanism of mulberry pistillate flower de- velopment.展开更多
AIM: To assess the proteome of normal versus tumor tissue in squamous cell carcinoma of the esophagus (SCCE) in Iranian patients and compare our results with former reports by using proteomics. METHODS: Protein wa...AIM: To assess the proteome of normal versus tumor tissue in squamous cell carcinoma of the esophagus (SCCE) in Iranian patients and compare our results with former reports by using proteomics. METHODS: Protein was extracted from normal and tumor tissues. Two dimensional electrophoresis was carried out and spots with differential expression were identified with mass spectrometry. RNA extraction and RT-PCR along with immunodetection were performed. RESULTS: Fourteen proteins were found whose expression levels differed in tumor compared to normal tissues. Mass spectrometric analysis resulted in the identification of β-tropomyosin (TMβ), myosin light chain 2 (and its isoform), myosin regulatory light chain 2, peroxyredoxin 2, annexin I and an unknown polypeptide as the down regulated polypeptides in tumor tissue. Heat shock protein 70 (HSP70), TPM4-ALK fusion oncoprotein 2, myosin light polypeptide 6, keratin I, GH16431p and calreticulin were the up-regulated polypeptides found in tumor tissue. Several of these proteins, such as TMβ, HSP70, annexin Ⅰ, calreticulin, TPM4-ALK and isoforms of myosins, have been well recognized in tumorigenesis of esophageal or other types of cancers. CONCLUSION: Our study not only supports the involvement of some of the formerly reported proteins in SCCE but also introduces additional proteins found to be lost in SCCE, including TMβ.展开更多
Aim To analyze the secondary structure and neurotrophic effect of a specific protein in sensory neurons. Methods Comparison of the proteins expressed in the rat spinal sensory neurons and motor neurons was made by t...Aim To analyze the secondary structure and neurotrophic effect of a specific protein in sensory neurons. Methods Comparison of the proteins expressed in the rat spinal sensory neurons and motor neurons was made by two dimensional electrophoresis. One specific protein in sensory neurons was isolated and purified by DEAE Sephacel ion exchange chromatography and high performance liquid chromatography. A primary analysis of its secondary structure by circular dichroism, and its neurotrophic effects were investigated using the model of dorsal root ganglia(DRG) cultured in vitro . Results The molecular weight and isoelectric point of the protein were 33 1 kDa and 5 52, respectively. Its circular dichroism showed that there were 20 8% α helix, 54 8% β sheet, 7 3% turn, and 17 1% random coil in its secondary structure. Biological experiments showed that the protein could promote the neurite outgrowth of DRG. Conclusion A specific protein in spinal sensory tissue with molecular weight of 33 1 kDa has been purified. There is mainly β sheet in the secondary structure of the protein. And the protein has neurotrophic effects in the model of DRG.展开更多
A new morphological variant strain of microsporidium was produced by infecting the mulberry looper, Hemerophilaatrilineata [Phthonandria atrilineata], with Nosema bombycis successively for 24 times, and named 24Nbh. C...A new morphological variant strain of microsporidium was produced by infecting the mulberry looper, Hemerophilaatrilineata [Phthonandria atrilineata], with Nosema bombycis successively for 24 times, and named 24Nbh. Comparativestudies on morphology, infectivity and spore surface protein were conducted. 24Nbh was short and wide, and had asignificant difference (P<0.01) over the Nb spores. The infectivity tests conducted on second instar silkworm larvaeshowed that IC50 of 24Nbh was 1.98104 spores mL-1 and of Nb was 1.72103 spores mL-1, thus indicating that the infectivityof Nb decreased 11.5 times after multiplying in mulberry looper for 24 times. The IC50 of spores from silkworm infected with24 Nbh was 6.9 times less than Nb, showing that the infectivity of 24Nbh spores rejuvenated very fast when reinfected tosilkworms, further more, the length and width of such spore was larger than 24Nbh (P<0.01) and smaller than Nb (P<0.05).The SDS-PAGE profiles of Nb and 24Nbh were generally the same, 4 distinct proteins of 12, 17, 30, 33 kDa were obtainedwith difference in quantity. When 120 g of protein was applied for 2D-PAGE, five suspected different proteins withdifference in quantity were observed. These results demonstrate that these differential proteins maybe associated withvariation in infectivity of the spores.展开更多
The differentially expressed proteins between normal and tamoxifen-induced autophagic MCF-7 cells were studied by 2-DE combining with mass spectrometric analysis.Totally,there were 668±99 and 550±97 protein ...The differentially expressed proteins between normal and tamoxifen-induced autophagic MCF-7 cells were studied by 2-DE combining with mass spectrometric analysis.Totally,there were 668±99 and 550±97 protein spots detected in the 2-DE maps from normal and autophagic MCF-7 cells respectively,among which 5 changed protein spots were identified by the Q-TOF MS/MS and database search.展开更多
Arsenic(As)is broadly distributed due to natural and anthropogenic sources,and it is toxic to organisms.This study aimed to investigate the proteomic response in earthworm exposed to As^(3+).Earthworms were exposed to...Arsenic(As)is broadly distributed due to natural and anthropogenic sources,and it is toxic to organisms.This study aimed to investigate the proteomic response in earthworm exposed to As^(3+).Earthworms were exposed to As^(3+)in soil at 5-80 mg kg1,and samples were collected after 60 days exposure.Two-dimensional electrophoresis(2-DE)was used to separate the proteins in earthworm homogenate,then differentially expressed proteins(DEPs)were identified using MALDI-TOF/TOF-MS analysis.After 2-DE,36 DEPs were found and 24 of them were successfully identified.79.2%of DEPs were upregulated compared to the control group.The maximum fold change reached 53.8 in spot 3108 in the 80 mg kg^(-1)As group.Two proteins were not found in the control group but found in the As treated groups.Proteins were grouped into metabolism,signal transduction,stress-related,transport,regulation,and predicted/hypothetical protein categories based on their function.The protein-protein interaction between the DEPs indicated that serum albumin(ALB)is very important,related to 6 other proteins.Proteins were then verified by western blot,the results were in agreement with the proteomic analyses.The identification of induced or repressed proteins because of As^(3+)in earthworms is helpful to explore the underlying mechanisms of soil arsenic ecotoxicity.展开更多
Background Equatorial lens epithelial cells proliferate and differentiate into fiber cells throughout life, while central lens epithelial cells proliferate little and do not form fiber ceils. This study aimed to inves...Background Equatorial lens epithelial cells proliferate and differentiate into fiber cells throughout life, while central lens epithelial cells proliferate little and do not form fiber ceils. This study aimed to investigate the differences in gene expression between the central and the peripheral epithelial cells of the bovine lens. Methods Lens epithelia were dissected into central (≤11.5 mm diameter, cLEC) and peripheral regions (pLEC). The differences in gene expression and protein accumulation between these two regions were assayed by microarray analysis and two-dimensional electrophoresis (2-DE) combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Differently expressed proteins were validated by immunoanalyses. Results By microarray analysis, 67 transcripts were at least two-fold lower and 269 at least two-fold higher in pLEC compared with that in cLEC. Thirty-four protein spots, including 20 in cLEC and 14 in pLEC, were identified by two dimensional electrophoresis and mass spectrometry. Of these 34 protein products, 28 were represented by probe sets on the microarray. Nine transcripts changed in the same direction and four transcripts in the opposite direction to their protein products. Immunoanalyses revealed that three (mitogen-activated protein kinase 1 (MAPK1), nidogen (NID), small nuclear ribonucleoprotein N (SNRPN)) out of four transcripts with opposite change between 2-DE and microarray assay showed the same changes as the results of 2-DE gel analyses. The genes differently expressed between cLEC and pLEC mainly include those related to the MAPK, transforming growth factor β (TGFβ) signaling and glycolysis pathways. Conclusion The results suggested that there were distinctly different genome activities, including a specific group of pathways, between central and peripheral lens epithelial cells.展开更多
To develop an economical and feasible approach to probe protein complexes, differential centrifugation and three-dimensional polyacrylamide gel electrophoresis (PAGE) were performed to separate protein complexes fro...To develop an economical and feasible approach to probe protein complexes, differential centrifugation and three-dimensional polyacrylamide gel electrophoresis (PAGE) were performed to separate protein complexes from the cell lysate of human pancreatic cancer cell line, SW1990, followed by mass spectrometric identification. Four macromolecular protein complexes were separated and identified unambiguously.展开更多
Proteomics involves the separation of proteins,identification of the amino acid sequence of the interested or target proteins,study of the function of the proteins,modification,structure and ultimate assignments to fu...Proteomics involves the separation of proteins,identification of the amino acid sequence of the interested or target proteins,study of the function of the proteins,modification,structure and ultimate assignments to functional pathways in the cell.The proteomic investigations have contributed greatly to human diseases studies,new drugs discovery researches,and environmental science in recent years.This article provides a review on the development of the main proteomic technologies,including both the gel based and non-gel based technologies,and their applications in environmental science.Proteomic technologies have been utilized in the environmental stresses studies to analyze the induction or reduction of proteins at expression level and identify the target proteins to investigate their function in response to environmental stresses,such as high or low pH,oxidation stress,and toxic chemicals.Such protein responses are also helpful to understand the mechanisms of some cellular activities and the functions of some proteins.展开更多
基金supported by The General Program of National Natural Science Foundation of China(81071314)
文摘Objective To investigate the protein expression profiles of the major food‐borne pathogen Campylobacter jejuni NCTC11168.Methods Membrane and soluble cellular proteins were extracted from the genome‐sequenced C.jejuni strain NCTC11168.Protein expression profiles were determined using two‐dimensional gel electrophoresis(2‐DE).All the detected spots on the 2‐DE map were subjected to matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry(MALDI‐TOF/TOF) analysis.Results A total of 537 and 333 spots were detected from the whole cell and membrane‐associated proteins of C.jejuni NCTC11168 cultured on Columbia agar medium at 42 ℃ by 2‐DE and Coomassie Brilliant Blue staining,respectively.Analyses of whole cell and membrane‐associated proteins included 399 and 133 spots,respectively,which included 182 and 53 functional proteins identified by MALDI‐TOF/TOF analysis.Conclusion The comprehensive expression protein profiles of C.jeuni NCTC11168 obtained in this study will be useful for elucidating the roles of these proteins in further pathogenesis investigation.
基金Supported by National Natural Science Foundation of China(31072087)~~
文摘[Objective] This study aimed to explore the proteins related to pistillate flower development in different mulberry cultivars. [Method] The total proteins of the pistillate flowers of two mulberry cultivars Dal0 (Morus atropurpurea Roxb.) and SG01 (Morus muIticaulis Perr.) were extracted, separated and detected through two- dimensional electrophoresis (2-DE) and mass spectrometry. [Result] There was sig- nificant difference in the expression of proteins from the pistillate flowers of different mulberry cultivars. From the 2-DE images of Dal0 and SG01, 445_+17 and 425_+12 protein spots were respectively detected. The expression levels of 75 protein spots differed significantly. Thirteen spots those were expressed at high levels and well separated were analyzed by mass spectrometry, and nine of them were identified successfully. The nine proteins are involved in the glycometabolism, protein and amino acid metabolism and defense responses during the development of mulberry pistillate flower after they were pollinated. [Conclusion] The findings will provide reference for further study on the molecular mechanism of mulberry pistillate flower de- velopment.
基金Supported by National Institute of Genetic Engineering and Biotechnology, No. 199 and Digestive Disease Research Center, No. 18/81
文摘AIM: To assess the proteome of normal versus tumor tissue in squamous cell carcinoma of the esophagus (SCCE) in Iranian patients and compare our results with former reports by using proteomics. METHODS: Protein was extracted from normal and tumor tissues. Two dimensional electrophoresis was carried out and spots with differential expression were identified with mass spectrometry. RNA extraction and RT-PCR along with immunodetection were performed. RESULTS: Fourteen proteins were found whose expression levels differed in tumor compared to normal tissues. Mass spectrometric analysis resulted in the identification of β-tropomyosin (TMβ), myosin light chain 2 (and its isoform), myosin regulatory light chain 2, peroxyredoxin 2, annexin I and an unknown polypeptide as the down regulated polypeptides in tumor tissue. Heat shock protein 70 (HSP70), TPM4-ALK fusion oncoprotein 2, myosin light polypeptide 6, keratin I, GH16431p and calreticulin were the up-regulated polypeptides found in tumor tissue. Several of these proteins, such as TMβ, HSP70, annexin Ⅰ, calreticulin, TPM4-ALK and isoforms of myosins, have been well recognized in tumorigenesis of esophageal or other types of cancers. CONCLUSION: Our study not only supports the involvement of some of the formerly reported proteins in SCCE but also introduces additional proteins found to be lost in SCCE, including TMβ.
文摘Aim To analyze the secondary structure and neurotrophic effect of a specific protein in sensory neurons. Methods Comparison of the proteins expressed in the rat spinal sensory neurons and motor neurons was made by two dimensional electrophoresis. One specific protein in sensory neurons was isolated and purified by DEAE Sephacel ion exchange chromatography and high performance liquid chromatography. A primary analysis of its secondary structure by circular dichroism, and its neurotrophic effects were investigated using the model of dorsal root ganglia(DRG) cultured in vitro . Results The molecular weight and isoelectric point of the protein were 33 1 kDa and 5 52, respectively. Its circular dichroism showed that there were 20 8% α helix, 54 8% β sheet, 7 3% turn, and 17 1% random coil in its secondary structure. Biological experiments showed that the protein could promote the neurite outgrowth of DRG. Conclusion A specific protein in spinal sensory tissue with molecular weight of 33 1 kDa has been purified. There is mainly β sheet in the secondary structure of the protein. And the protein has neurotrophic effects in the model of DRG.
基金funded by the National Natural Science Foundation of China(30270898).
文摘A new morphological variant strain of microsporidium was produced by infecting the mulberry looper, Hemerophilaatrilineata [Phthonandria atrilineata], with Nosema bombycis successively for 24 times, and named 24Nbh. Comparativestudies on morphology, infectivity and spore surface protein were conducted. 24Nbh was short and wide, and had asignificant difference (P<0.01) over the Nb spores. The infectivity tests conducted on second instar silkworm larvaeshowed that IC50 of 24Nbh was 1.98104 spores mL-1 and of Nb was 1.72103 spores mL-1, thus indicating that the infectivityof Nb decreased 11.5 times after multiplying in mulberry looper for 24 times. The IC50 of spores from silkworm infected with24 Nbh was 6.9 times less than Nb, showing that the infectivity of 24Nbh spores rejuvenated very fast when reinfected tosilkworms, further more, the length and width of such spore was larger than 24Nbh (P<0.01) and smaller than Nb (P<0.05).The SDS-PAGE profiles of Nb and 24Nbh were generally the same, 4 distinct proteins of 12, 17, 30, 33 kDa were obtainedwith difference in quantity. When 120 g of protein was applied for 2D-PAGE, five suspected different proteins withdifference in quantity were observed. These results demonstrate that these differential proteins maybe associated withvariation in infectivity of the spores.
文摘The differentially expressed proteins between normal and tamoxifen-induced autophagic MCF-7 cells were studied by 2-DE combining with mass spectrometric analysis.Totally,there were 668±99 and 550±97 protein spots detected in the 2-DE maps from normal and autophagic MCF-7 cells respectively,among which 5 changed protein spots were identified by the Q-TOF MS/MS and database search.
基金supported by the Shanghai Agriculture Applied Technology Development Program of China(2021 No.2-2)the Natural Science Projects of Henan University of Technology,China(No.2019BS037)+1 种基金the Open Fund Project of State Environmental Protection Key Laboratory of Monitoring for Heavy Metal Pollutants(SKLMHM202106)the National Natural Science Foundation of China(No.41471203).
文摘Arsenic(As)is broadly distributed due to natural and anthropogenic sources,and it is toxic to organisms.This study aimed to investigate the proteomic response in earthworm exposed to As^(3+).Earthworms were exposed to As^(3+)in soil at 5-80 mg kg1,and samples were collected after 60 days exposure.Two-dimensional electrophoresis(2-DE)was used to separate the proteins in earthworm homogenate,then differentially expressed proteins(DEPs)were identified using MALDI-TOF/TOF-MS analysis.After 2-DE,36 DEPs were found and 24 of them were successfully identified.79.2%of DEPs were upregulated compared to the control group.The maximum fold change reached 53.8 in spot 3108 in the 80 mg kg^(-1)As group.Two proteins were not found in the control group but found in the As treated groups.Proteins were grouped into metabolism,signal transduction,stress-related,transport,regulation,and predicted/hypothetical protein categories based on their function.The protein-protein interaction between the DEPs indicated that serum albumin(ALB)is very important,related to 6 other proteins.Proteins were then verified by western blot,the results were in agreement with the proteomic analyses.The identification of induced or repressed proteins because of As^(3+)in earthworms is helpful to explore the underlying mechanisms of soil arsenic ecotoxicity.
文摘Background Equatorial lens epithelial cells proliferate and differentiate into fiber cells throughout life, while central lens epithelial cells proliferate little and do not form fiber ceils. This study aimed to investigate the differences in gene expression between the central and the peripheral epithelial cells of the bovine lens. Methods Lens epithelia were dissected into central (≤11.5 mm diameter, cLEC) and peripheral regions (pLEC). The differences in gene expression and protein accumulation between these two regions were assayed by microarray analysis and two-dimensional electrophoresis (2-DE) combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Differently expressed proteins were validated by immunoanalyses. Results By microarray analysis, 67 transcripts were at least two-fold lower and 269 at least two-fold higher in pLEC compared with that in cLEC. Thirty-four protein spots, including 20 in cLEC and 14 in pLEC, were identified by two dimensional electrophoresis and mass spectrometry. Of these 34 protein products, 28 were represented by probe sets on the microarray. Nine transcripts changed in the same direction and four transcripts in the opposite direction to their protein products. Immunoanalyses revealed that three (mitogen-activated protein kinase 1 (MAPK1), nidogen (NID), small nuclear ribonucleoprotein N (SNRPN)) out of four transcripts with opposite change between 2-DE and microarray assay showed the same changes as the results of 2-DE gel analyses. The genes differently expressed between cLEC and pLEC mainly include those related to the MAPK, transforming growth factor β (TGFβ) signaling and glycolysis pathways. Conclusion The results suggested that there were distinctly different genome activities, including a specific group of pathways, between central and peripheral lens epithelial cells.
基金Project supported by the National High-Tech R&D Program of China (863 Program, No. 2006AA02Z154) and the National Natural Science Foundation of China (No. 21075 137).
文摘To develop an economical and feasible approach to probe protein complexes, differential centrifugation and three-dimensional polyacrylamide gel electrophoresis (PAGE) were performed to separate protein complexes from the cell lysate of human pancreatic cancer cell line, SW1990, followed by mass spectrometric identification. Four macromolecular protein complexes were separated and identified unambiguously.
文摘Proteomics involves the separation of proteins,identification of the amino acid sequence of the interested or target proteins,study of the function of the proteins,modification,structure and ultimate assignments to functional pathways in the cell.The proteomic investigations have contributed greatly to human diseases studies,new drugs discovery researches,and environmental science in recent years.This article provides a review on the development of the main proteomic technologies,including both the gel based and non-gel based technologies,and their applications in environmental science.Proteomic technologies have been utilized in the environmental stresses studies to analyze the induction or reduction of proteins at expression level and identify the target proteins to investigate their function in response to environmental stresses,such as high or low pH,oxidation stress,and toxic chemicals.Such protein responses are also helpful to understand the mechanisms of some cellular activities and the functions of some proteins.