scholarly journals Study on Differential Protein Expression in Natural Selenium-Enriched and Non-Selenium-Enriched Rice Based on iTRAQ Quantitative Proteomics

Biomolecules ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 130 ◽  
Author(s):  
Rui Zeng ◽  
Muhammad Farooq ◽  
Li Wang ◽  
Yang Su ◽  
Tengda Zheng ◽  
...  
Keyword(s):  
Amino Acid ◽  
Differential Expression ◽  
Starch Synthesis ◽  
Oxygen Metabolism ◽  
Absolute Quantification ◽  
Stress Effect ◽  
Cysteine Synthase ◽  
Liquid Chromatography Tandem Mass ◽  
Isobaric Tags ◽  
Shock Proteins

This work was designated to scrutinize the protein differential expression in natural selenium-enriched and non-selenium-enriched rice using the Isobaric-tags for relative and absolute quantification (iTRAQ) proteomics approach. The extracted proteins were subjected to enzyme digestion, desalting, and identified by iTRAQ coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology. High pH C18 separation analysis was performed, and the data were then analyzed by Protein PilotTM (V4.5) search engine. Protein differential expression was searched out by comparing relatively quantified proteins. The analysis was conducted using gene ontology (GO), cluster of orthologous groups of proteins (COG) and Kyoto encyclopedia of genes and genomes (KEGG) metabolic pathways. A total of 3235 proteins were detected and 3161 proteins were quantified, of which 401 were differential proteins. 208 down-regulated and 193 up-regulated proteins were unveiled. 77 targeted significant differentially expressed proteins were screened out for further analysis, and were classified into 10 categories: oxidoreductases, transferases, isomerases, heat shock proteins, lyases, hydrolases, ligases, synthetases, tubulin, and actin. The results indicated that the anti-stress, anti-oxidation, active oxygen metabolism, carbohydrate and amino acid metabolism of natural selenium-enriched rice was higher than that of non-selenium rice. The activation of the starch synthesis pathway was found to be bounteous in non-selenium-enriched rice. Cysteine synthase (CYS) and methyltransferase (metE) might be the two key proteins that cause amino acid differences. OsAPx02, CatC, riPHGPX, HSP70 and HSP90 might be the key enzymes regulating antioxidant and anti-stress effect differences in two types of rice. This study provides basic information about deviations in protein mechanism and secondary metabolites in selenium-enriched and non-selenium-enriched rice.

scholarly journals Understanding the Toxin Effects of β-Zearalenol and HT-2 on Bovine Granulosa Cells Using iTRAQ-Based Proteomics

Animals ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 130
Author(s):  
Lian Li ◽  
Min Yang ◽  
Chengmin Li ◽  
Fangxiao Yang ◽  
Genlin Wang
Keyword(s):  
Granulosa Cells ◽  
Absolute Quantification ◽  
P Value ◽  
Toxic Response ◽  
Human Food Chain ◽  
Response Profile ◽  
Isobaric Tags ◽  
Shock Proteins ◽  
Cellular Components ◽  
Marker Molecule

Zearalenone (ZEA) and T-2 are the most common mycotoxins in grains and can enter the animal and human food-chain and cause many health disorders. To elucidate the toxic response profile, we stimulated bovine granulosa cells (GCs) with β-zearalenol or HT-2. Using isobaric tags for relative and absolute quantification (iTRAQ)-based proteomic, 178 and 291 differentially expressed proteins (DEPs, fold change ≥ 1.3 and p-value < 0.05) in β-zearalenol and HT-2 groups were identified, respectively. Among these DEPs, there were 66 common DEPs between β-zearalenol and HT-2 groups. These 66 DEPs were associated with 23 biological processes terms, 14 molecular functions terms, and 19 cellular components terms. Most heat shock proteins (HSPs) were involved in the toxic response. Reactive oxygen species accumulation, the endoplasmic reticulum (ER)-stress related marker molecule (GRP78), and apoptosis were activated. β-zearalenol and HT-2 inhibited oestradiol (E2) production. These results emphasized the important function of HSPs, clarified oxidative stress, and demonstrated the caspase-3 signaling cascade involved in mycotoxin-treated toxic response, along with decreased E2 production. This study offers new insights into the toxicity of β-zearalenol and HT-2 on ovarian granulosa cells.

scholarly journals Suckling Piglet Intestinal Enterocyte Nutrient Metabolism Changes

2018 ◽  
Vol 48 (5) ◽  
pp. 2103-2113 ◽  
Author(s):  
Qiye Wang ◽  
Xia Xiong ◽  
Xiaocheng Wang ◽  
Qiang Tu ◽  
Jianzhong Li ◽  
...  
Keyword(s):  
Amino Acids ◽  
Fatty Acids ◽  
Absolute Quantification ◽  
Intestinal Morphology ◽  
Nutrient Metabolism ◽  
Biological Regulation ◽  
Citrate Cycle ◽  
Cellular Processes ◽  
Liquid Chromatography Tandem Mass ◽  
Isobaric Tags

Background/Aims: Intestinal morphology and the types of enterocytes are changed in piglets during the suckling period, but it is unclear whether these changes are associated with metabolic changes in epithelium. The present study was conducted to test the hypothesis that glucose, fatty acids, and amino acid metabolism in differentiated piglet enterocytes changed during suckling. Methods: Twenty-four piglets (Duroc × [Landrace × Yorkshire]) from 8 litters (3 piglets/litter) were selected. A single piglet from each litter was randomly selected and euthanized at days 7, 14, and 21. Differentiated enterocytes (DE) were isolated from their mid-jejunum. Isobaric tags for relative and absolute quantification and subsequent liquid chromatography-tandem mass spectrometry were used to identify and measure protein synthesis. Results: The results showed that various activities, including: cellular processes; metabolic processes; biological regulation; pigmentation; and, localization, in DEs changed during suckling. Metabolic process analyses revealed that protein expression related to glycolysis and citrate cycle was decreased from day 7 to day 14. The number of differentiated enterocytes of 21 d piglets increased compared to 7 d piglets. Most of the proteins involved in fatty acid and amino acids metabolism had decreased DE expression between day 7 and day 14. Some, but not all, detected proteins down-regulated in DEs of 21 day piglets compared to 7 day piglets. Conclusion: These results indicate that glucose, fatty acids, and amino acids metabolism changed during suckling. This may provide useful information for designing feed formulas and regulating piglet intestinal growth and development.

iTRAQ-Based Quantitative Proteomic Analysis of Pseudostellaria heterophylla from Geo-Authentic Habitat and Cultivated Bases

2019 ◽  
Vol 16 (3) ◽  
pp. 231-245
Author(s):  
Yujiao Hua ◽  
Chengcheng Wang ◽  
Shengnan Wang ◽  
Zixiu Liu ◽  
Xunhong Liu ◽  
...  
Keyword(s):  
Binding Proteins ◽  
Large Subunit ◽  
Absolute Quantification ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Realtime Pcr ◽  
Pseudostellaria Heterophylla ◽  
Response To Stress ◽  
Isobaric Tags ◽  
Shock Proteins

Background: Pseudostellaria heterophylla is an important tonic traditional Chinese medicine. However, the molecular changes in the herb from geo-authentic habitat and cultivated bases remain to be explored. Objective: The purpose of this research was to study differences in P. heterophylla from geo-authentic habitat and cultivated bases. Methods: High-throughput technologies of transcriptomic and proteomic were used to identify proteins. Isobaric Tags for Relative and Absolute Quantification (iTRAQ) MS/MS has been utilized to evaluate changes in P. heterophylla from geo-authentic habitat and cultivated bases. Results: In this study, a total of 3775 proteins were detected, and 140 differentially expressed proteins were found in P. heterophylla from geo-authentic habitat and cultivated bases. 44 significantly differential expressed proteins were identified based on functional analysis classified into nine categories. Five differentially expressed proteins were confirmed at the gene expression level by Quantitative realtime PCR. Catabolic metabolism, carbohydrate metabolism, and response to stress of oxidoreductases and transferases in P. heterophylla from geo-authentic habitat were stronger than in those from cultivated bases, but protein folding and response to stress of heat shock proteins, isomerases, rubisco large subunit-binding proteins, chaperone proteins, and luminal-binding proteins in herbs from cultivated bases were more active. ADG1 and TKTA could be the critical proteins to regulate sucrose; MFP2 and CYS may be the crucial proteins that control the metabolism of fatty acids and amino acids. Conclusion: These results will provide the basic information for exploring the differences in secondary metabolites in P. heterophylla from geo-authentic habitat and cultivated bases and the protein mechanism of its quality formation.

scholarly journals iTRAQ-based Quantitative Proteomics Analysis Identifies Host Pathways Modulated during Toxoplasma gondii Infection in Swine

2020 ◽  
Vol 8 (4) ◽  
pp. 518 ◽  
Author(s):  
Jun-Jun He ◽  
Jun Ma ◽  
Jin-Lei Wang ◽  
Fu-Kai Zhang ◽  
Jie-Xi Li ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Protein Expression ◽  
Absolute Quantification ◽  
Mesenteric Lymph Nodes ◽  
Proteomics Analysis ◽  
Liquid Chromatography Tandem Mass ◽  
Isobaric Tags ◽  
Toxoplasma Gondii Infection ◽  
The Brain

Toxoplasma gondii is a leading cause of foodborne illness and consumption of undercooked pig meat is a major risk factor for acquiring toxoplasmosis, which causes a substantial burden on society. Here, we used isobaric tags for relative and absolute quantification (iTRAQ) labelling coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify cellular proteins and pathways altered during T. gondii infection in pigs. We also used parallel reaction monitoring-based LC-MS/MS to verify the levels of protein expression of infected spleens and mesenteric lymph nodes (MLNs). At 6 days post-infection (dpi), 156, 391, 170, 292, and 200 differentially expressed proteins (DEPs) were detected in the brain, liver, lung, MLNs and spleen, respectively. At 18 dpi, 339, 351, 483, 388, and 303 DEPs were detected in the brain, liver, lung, MLNs and spleen, respectively. Although proteins involved in immune responses were upregulated in all infected tissues, protein expression signature in infected livers was dominated by downregulation of the metabolic processes. By weighted gene co-expression network analysis, we could further show that all proteins were clustered into 25 co-expression modules and that the pink module significantly correlated with the infection status. We also identified 163 potential anti-T. gondii proteins (PATPs) and provided evidence that two PATPs (HSP70.2 and PDIA3) can reduce T. gondii burden in porcine macrophages in vitro. This comprehensive proteomics analysis reveals new facets in the pathogenesis of T. gondii infection and identifies key proteins that may contribute to the pig’s defense against this infection.

scholarly journals Study on inhibitory effect of cinnamaldehyde on Salmonella based on iTRAQ technology

2020 ◽  
Author(s):  
Lizi Yin ◽  
Han Chen ◽  
Xuewen He ◽  
Zhongqiong Yin ◽  
Hualin Fu ◽  
...  
Keyword(s):  
Salmonella Typhimurium ◽  
Absolute Quantification ◽  
Inhibitory Effect ◽  
Kegg Pathways ◽  
Therapeutic Drugs ◽  
Liquid Chromatography Tandem Mass ◽  
New Ideas ◽  
Isobaric Tags ◽  
Novel Drug ◽  
Cellular Components

Abstract BackgroundSalmonella is one of the most concerned pathogenic bacterium worldwide. In our previous experiments, cinnamaldehyde showed prominent antibacterial effects on Salmonella Typhimurium (S. Typhimurium), indicating that it could be developed as a novel therapeutic drugs for Salmonella.ResultsIn this assay, the mechanism of cinnamaldehyde inhibiting S. Typhimurium was investigated. We studied the antibacterial mechanism of cinnamaldehyde using isobaric tags for relative and absolute quantification (iTRAQ) with two-dimensional liquid chromatography/tandem mass spectrometry (2D-LC-MS/MS), combining with bioinformatics. There were 2,212 proteins detected by iTRAQ, of which 73 proteins were up-regulated, and 82 of proteins were down-regulated. The differently expressed proteins were connected with 10 cellular components, 9 molecular functions and 13 biological processes as well as 57 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. There were some differently expressed proteins involved into biosynthesis and metabolism of amino, redox reaction, energy metabolism, toxins and drug-resistance. Cinnamaldehyde may have the potential to become a novel drug for Salmonella infection.ConclusionsBased on the iTRAQ technology, this paper explores the mechanism of cinnamaldehyde on Salmonella typhimurium at the protein level, digs the target of action, lays a theoretical foundation for further exploration, and provides new ideas for the clinical use and development of cinnamaldehyde.Therefore, cinnamaldehyde may have the potential to be used combined with antibiotics.

scholarly journals Isobaric Tags for Relative and Absolute Quantification-Based Proteomics Reveals Candidate Proteins of Fat Deposition in Chinese Indigenous Sheep With Morphologically Different Tails

2021 ◽  
Vol 12 ◽  
Author(s):  
Caiye Zhu ◽  
Heping Cheng ◽  
Na Li ◽  
Tiaoguo Liu ◽  
Youji Ma
Keyword(s):  
Absolute Quantification ◽  
Scientific Basis ◽  
Phenotypic Differences ◽  
Kegg Pathways ◽  
Indigenous Sheep ◽  
Genes Encoding ◽  
Liquid Chromatography Tandem Mass ◽  
Isobaric Tags ◽  
Function Regulator ◽  
Pathway Analyses

Background: Chinese indigenous sheep can be classified into two types according to their tail morphology: fat-rumped and thin-tailed sheep, of which the typical breeds are Altay sheep and Tibetan sheep, respectively.Methods: To identify the differentially expressed proteins (DEPs) underlying the phenotypic differences between tail types, we used isobaric tags for relative and absolute quantification (iTRAQ) combined with multi-dimensional liquid chromatography tandem-mass spectrometry (LC-MS/MS) technology to detect candidate proteins. We then subjected these to a database search and identified the DEPs. Finally, bioinformatics technology was used to carry out Gene Ontology (GO) functional and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses.Results: A total of 3,248 proteins were identified, of which 44 were up-regulated and 40 were down-regulated DEPs. Analyzing their GO function terms and KEGG pathways revealed that the functions of these DEPs are mainly binding, catalytic activity, structural molecule activity, molecular function regulator, and transporter activity. Among the genes encoding the DEPs, APOA2, GALK1, ADIPOQ, and NDUFS4 are associated with fat formation and metabolism.Conclusion: The APOA2, GALK1, ADIPOQ, and NDUFS4 genes may be involved in the deposition of fat in the tail of sheep. This study provides a scientific basis for the breeding of thin-tailed sheep.

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