scholarly journals Proteomic Analyses of Thioredoxins f and m Arabidopsis thaliana Mutants Indicate Specific Functions for These Proteins in Plants

Antioxidants ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 54 ◽  
Author(s):  
Juan Fernández-Trijueque ◽  
Antonio-Jesús Serrato ◽  
Mariam Sahrawy
Keyword(s):  
Protein Profiling ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Specific Target ◽  
Fructose 1,6 Bisphosphatase ◽  
Response To Stress ◽  
Quantitative Alteration ◽  
Hormone Signalling ◽  
Specific Target Protein

A large number of plastidial thioredoxins (TRX) are present in chloroplast and the specificity versus the redundancy of their functions is currently under discussion. Several results have highlighted the fact that each TRX has a specific target protein and thus a specific function. In this study we have found that in vitro activation of the fructose-1,6-bisphosphatase (FBPase) enzyme is more efficient when f1 and f2 type thioredoxins (TRXs) are used, whilst the m3 type TRX did not have any effect. In addition, we have carried out a two-dimensional electrophoresis-gel to obtain the protein profiling analyses of the trxf1, f2, m1, m2, m3 and m4 Arabidopsis mutants. The results revealed quantitative alteration of 86 proteins and demonstrated that the lack of both the f and m type thioredoxins have diverse effects on the proteome. Interestingly, 68% of the differentially expressed proteins in trxf1 and trxf2 mutants were downregulated, whilst 75% were upregulated in trxm1, trxm2, trxm3 and trxm4 lines. The lack of TRX f1 provoked a higher number of down regulated proteins. The contrary occurred when TRX m4 was absent. Most of the differentially expressed proteins fell into the categories of metabolic processes, the Calvin–Benson cycle, photosynthesis, response to stress, hormone signalling and protein turnover. Photosynthesis, the Calvin–Benson cycle and carbon metabolism are the most affected processes. Notably, a significant set of proteins related to the answer to stress situations and hormone signalling were affected. Despite some studies being necessary to find specific target proteins, these results show signs that are suggest that the f and m type plastidial TRXs most likely have some additional specific functions.

scholarly journals Dysregulation of ferroptosis may involve in the development of non-small cell lung cancer in Xuanwei area

2020 ◽  
Author(s):  
Zhang Yang ◽  
Guangjian Li ◽  
Jiapeng Yang ◽  
Guangqiang Zhao ◽  
Zhenghai Shen ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Indoor Air Pollution ◽  
Protein Profiling ◽  
Differentially Expressed ◽  
Receptor Interaction ◽  
Differentially Expressed Proteins ◽  
Tandem Mass Tag ◽  
Differential Proteins ◽  
Local Residents

Abstract Background The Xuanwei area of Yunnan Province, China is one of the regions with the highest incidence of lung cancer in the world. Local residents use bituminous coal as fuel for cooking and heating, which causes serious indoor air pollution. After the local government carried out furnace and stove reform work, the high incidence of lung cancer in residents continued. We herein wonder if there are specific mechanisms at protein level for the development of lung cancer in the area. Methods We investigated the changes of protein profiling in tumor of the patients from Xuanwei area. Tandem Mass Tag (TMT) were employed to screen the differential proteins between carcinoma and para-carcinoma tissues. Results We identified a total of 422 differentially-expressed proteins, among which 162 proteins were significantly upregulated and 260 were downregulated compared to para-carcinoma tissues. Many of the differentially-expressed proteins were related to ECM-receptor interaction, focal adhesion, PI3K/AKT pathway and ferroptosis. Further experiments on the two differential proteins, TXN2 and HP, showed that the change of their expressions could make the lung cancer cell lines more resistant to erastin or RSL-induced ferroptosis in vitro, and promote the growth of tumor in nude mice. Conclusion This study revealed that aberrant regulation of ferroptosis may involve in the development of lung cancer in Xuanwei area.

In vitro andin silico processes to identify differentially expressed proteins

PROTEOMICS ◽  
2004 ◽  
Vol 4 (8) ◽  
pp. 2333-2351 ◽  
Author(s):  
Nadia Allet ◽  
Nicolas Barrillat ◽  
Thierry Baussant ◽  
Celia Boiteau ◽  
Paolo Botti ◽  
...  
Keyword(s):  
Differentially Expressed ◽  
Differentially Expressed Proteins

scholarly journals Two novel non-cationic defensin-like antimicrobial peptides from haemolymph of the female tick, Amblyomma hebraeum

2004 ◽  
Vol 379 (3) ◽  
pp. 681-685 ◽  
Author(s):  
Ren LAI ◽  
Lee O. LOMAS ◽  
Jan JONCZY ◽  
Philip C. TURNER ◽  
Huw H. REES
Keyword(s):  
Antimicrobial Peptides ◽  
Suppression Subtractive Hybridization ◽  
Subtractive Hybridization ◽  
Protein Profiling ◽  
Differentially Expressed ◽  
Cdna Clones ◽  
Hard Tick ◽  
Differentially Expressed Proteins ◽  
Amblyomma Hebraeum ◽  
Cdna Sequences

Two non-cationic defensin-like antimicrobial peptides, named Amblyomma defensin peptide 1 and Amblyomma defensin peptide 2, were identified from the hard tick, Amblyomma hebraeum, by a combination of suppression subtractive hybridization for differentially expressed genes and proteomics. cDNA clones encoding each of these two defensin-like antimicrobial peptides were isolated from the differentially expressed cDNA library of the tick synganglia (central nervous system). The preproproteins deduced from the cDNA sequences each have 92 amino acid residues. Amblyomma defensin peptide 2 was purified from the haemolymph of fed female ticks. The purified peptide displayed antibacterial activity against Gram-negative and Gram-positive bacteria. Amblyomma defensin peptide 1 was further identified by protein chip capture combined with SELDI-TOF (surface-enhanced laser desorption/ionization–time-of-flight) MS. By screening for differentially expressed proteins, it was found that the expression of Amblyomma defensin peptide 1 was upregulated during 4 days post-feeding. Our findings firstly provide two defensin-like antimicrobial peptides that are particularly novel in being anionic, together with corresponding cDNA sequences, in hard ticks, and prove that the combination of suppression subtractive hybridization and protein profiling is a powerful method to study differentially expressed proteins, especially for organisms without available genome sequence information.

scholarly journals Screening for proteins related to the biosynthesis of hispidin and its derivatives in Phellinus igniarius using iTRAQ proteomic analysis

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Jinjing Guo ◽  
Xiaoxi Liu ◽  
Yuanjie Li ◽  
Hongyan Ji ◽  
Cheng Liu ◽  
...  
Keyword(s):  
Stress Responses ◽  
Biosynthetic Pathway ◽  
Multiple Reaction Monitoring ◽  
Enrichment Analysis ◽  
Differentially Expressed ◽  
Common Source ◽  
Differentially Expressed Proteins ◽  
Phellinus Igniarius ◽  
Chemical Structures

Abstract Background Hispidin (HIP) and its derivatives, a class of natural fungal metabolites, possess complex chemical structures with extensive pharmacological activities. Phellinus igniarius, the most common source of HIP, can be used as both medicine and food. However, the biosynthetic pathway of HIP in P. igniarius remains unclear and we have a limited understanding of the regulatory mechanisms related to HIP. In this work, we sought to illustrate a biosynthesis system for hispidin and its derivatives at the protein level. Results We found that tricetolatone (TL) is a key biosynthetic precursor in the biosynthetic pathway of hispidin and that its addition led to increased production of hispidin and various hispidin derivatives. Based on the changes in the concentrations of precursors and intermediates, key timepoints in the biosynthetic process were identified. We used isobaric tags for relative and absolute quantification (iTRAQ) to study dynamic changes of related proteins in vitro. The 270 differentially expressed proteins were determined by GO enrichment analysis to be primarily related to energy metabolism, oxidative phosphorylation, and environmental stress responses after TL supplementation. The differentially expressed proteins were related to ATP synthase, NAD binding protein, oxidoreductase, and other elements associated with electron transfer and dehydrogenation reactions during the biosynthesis of hispidin and its derivatives. Multiple reaction monitoring (MRM) technology was used to selectively verify the iTRAQ results, leading us to screen 11 proteins that were predicted to be related to the biosynthesis pathways. Conclution These findings help to clarify the molecular mechanism of biosynthesis of hispidin and its derivatives and may serve as a foundation for future strategies to identify new hispidin derivatives.

scholarly journals Label-Free Quantitative Mass Spectrometry Reveals a Panel of Differentially Expressed Proteins in Colorectal Cancer

2015 ◽  
Vol 2015 ◽  
pp. 1-13 ◽  
Author(s):  
Nai-Jun Fan ◽  
Jiang-Ling Gao ◽  
Yan Liu ◽  
Wei Song ◽  
Zhan-Yang Zhang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Performance ◽  
Cell Structure ◽  
Protein Profiling ◽  
Differentially Expressed ◽  
Antimicrobial Protein ◽  
Differentially Expressed Proteins ◽  
Label Free ◽  
Mucosal Tissues ◽  
And Function

To identify potential biomarkers involved in CRC, a shotgun proteomic method was applied to identify soluble proteins in three CRCs and matched normal mucosal tissues using high-performance liquid chromatography and mass spectrometry. Label-free protein profiling of three CRCs and matched normal mucosal tissues were then conducted to quantify and compare proteins. Results showed that 67 of the 784 identified proteins were linked to CRC (28 upregulated and 39 downregulated). Gene Ontology and DAVID databases were searched to identify the location and function of differential proteins that were related to the biological processes of binding, cell structure, signal transduction, cell adhesion, and so on. Among the differentially expressed proteins, tropomyosin-3 (TPM3), endoplasmic reticulum resident protein 29 (ERp29), 18 kDa cationic antimicrobial protein (CAMP), and heat shock 70 kDa protein 8 (HSPA8) were verified to be upregulated in CRC tissue and seven cell lines through western blot analysis. Furthermore, the upregulation of TPM3, ERp29, CAMP, and HSPA8 was validated in 69 CRCs byimmunohistochemistry (IHC) analysis. Combination of TPM3, ERp29, CAMP, and HSPA8 can identify CRC from matched normal mucosal achieving an accuracy of 73.2% using IHC score. These results suggest that TPM3, ERp29, CAMP, and HSPA8 are great potential IHC diagnostic biomarkers for CRC.

scholarly journals Serum estradiol levels in controlled ovarian stimulation directly affect the endometrium

2017 ◽  
Vol 59 (2) ◽  
pp. 105-119 ◽  
Author(s):  
Kamran Ullah ◽  
Tanzil Ur Rahman ◽  
Hai-Tao Pan ◽  
Meng-Xi Guo ◽  
Xin-Yan Dong ◽  
...  
Keyword(s):  
Embryo Implantation ◽  
Controlled Ovarian Hyperstimulation ◽  
Endometrial Receptivity ◽  
Protein Profiling ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Serum Estradiol ◽  
Endometrial Cells ◽  
Ishikawa Cells ◽  
Estradiol Levels

Previous studies have shown that increasing estradiol concentrations had a toxic effect on the embryo and were deleterious to embryo adhesion. In this study, we evaluated the physiological impact of estradiol concentrations on endometrial cells to reveal that serum estradiol levels probably targeted the endometrium in controlled ovarian hyperstimulation (COH) protocols. An attachment model of human choriocarcinoma (JAr) cell spheroids to receptive-phase endometrial epithelial cells and Ishikawa cells treated with different estradiol (10−9 M or 10−7 M) concentrations was developed. Differentially expressed protein profiling of the Ishikawa cells was performed by proteomic analysis. Estradiol at 10−7 M demonstrated a high attachment rate of JAr spheroids to the endometrial cell monolayers. Using iTRAQ coupled with LC–MS/MS, we identified 45 differentially expressed proteins containing 43 significantly upregulated and 2 downregulated proteins in Ishikawa cells treated with 10−7 M estradiol. Differential expression of C3, plasminogen and kininogen-1 by Western blot confirmed the proteomic results. C3, plasminogen and kininogen-1 localization in human receptive endometrial luminal epithelium highlighted the key proteins as possible targets for endometrial receptivity and interception. Ingenuity pathway analysis of differentially expressed proteins exhibited a variety of signaling pathways, including LXR/RXR activation pathway and acute-phase response signaling and upstream regulators (TNF, IL6, Hmgn3 and miR-140-3p) associated with endometrial receptivity. The observed estrogenic effect on differential proteome dynamics in Ishikawa cells indicates that the human endometrium is the probable target for serum estradiol levels in COH cycles. The findings are also important for future functional studies with the identified proteins that may influence embryo implantation.

Relationship between seminal plasma tuberoinfundibular peptide of 39 residues and sperm functional attributes in buffalo (Bubalus bubalis)

2016 ◽  
Vol 28 (10) ◽  
pp. 1622 ◽  
Author(s):  
Sellappan Selvaraju ◽  
Lakshminarayana Somashekar ◽  
Binsila B. Krishnan ◽  
Sivashanmugam Parthipan ◽  
Guvvala Pushparani ◽  
...  
Keyword(s):  
Plasma Protein ◽  
Seminal Plasma ◽  
Sperm Quality ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Profiling ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Integrity Test ◽  
Tuberoinfundibular Peptide

The buffalo seminal plasma protein profile and its relationship with sperm quality have not been studied in detail. Thus, the aim of the present study was to profile buffalo seminal plasma proteins and to assess the relationship between differentially expressed proteins and sperm characteristics. Semen samples (n = 44) were collected from 11 Murrah buffalo bulls (four ejaculates from each animal) and seminal plasma protein profiling was performed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Matrix-assisted laser desorption ionisation time-of-flight analysis of one of the differentially expressed proteins, namely the 11–12 kDa protein, identified it as tuberoinfundibular peptide of 39 residues (TIP39). Western blot analysis confirmed the presence of TIP39, with TIP39 expression in seminal plasma varying among bulls. Based on TIP39 levels, bulls were classified into two groups, those with high and low protein. The percentages of spermatozoa positive for mitochondrial membrane potential test, chromatin distribution test, synthetic media sperm penetrability test and acrosomal integrity test were significantly (P < 0.05) high in the high protein group. The present study is the first to demonstrate the presence of TIP39 in buffalo seminal plasma and the positive effect of TIP39 on the functional parameters and fertilising ability of spermatozoa.

scholarly journals Proteomics variations after short-term heat shock treatment reveal differentially expressed proteins involved in early microspore embryogenesis in cabbage (Brassica oleracea)

2019 ◽  
Author(s):  
Henan Su ◽  
Guo Chen ◽  
Xing Liu ◽  
Limei Yang ◽  
Mu Zhuang ◽  
...  
Keyword(s):  
Heat Shock ◽  
In Vitro Culture ◽  
Microspore Embryogenesis ◽  
Shock Treatment ◽  
Differentially Expressed ◽  
High Temperature Treatment ◽  
Differentially Expressed Proteins ◽  
Heat Shock Treatment ◽  
Short Term

Abstract Background Microspore embryogenesis (ME) provides an efficient way to breed crops. In many cases, short-term heat shock treatment can greatly increase the embryogenesis rate of brassicas. However, its molecular mechanism is largely unclear. Results To mine for the key genes, pathways and interplay in the underlying networks, we compared the proteomes of isolated microspores with samples pre-treated at 32 °C for 24 h and 25 °C for 24 h using two cabbage accessions (Zhonggan 628 and 87-534) showing extremely different embryogenic rates. The embryo yield was 0 and 19.7 embryos/bud for Zhonggan 628 at 25 °C and 32 °C, respectively, and was 0 for 87-534 at both temperatures. Using a label-free proteomics technology, more differentially expressed proteins (DEPs) were found for Zhonggan 628 (363 DEPs, 115 upregulated and 248 downregulated) than for 87-534 (282 DEPs, 162 upregulated and 120 downregulated). 97 DEPs specially identified only in Zhonggan 628 but not in 87-534 after heat-shock treatment were the key proteins that maybe related to heat shock-induced embryogenesis in vitro culture. Those 97 DEPs were mainly enriched in carbon metabolic process and protein synthesis and degradation process. Malate dehydrogenase (mMDH), sgt1 homolog B (SGT1), heat shock 70 kDa protein 5 (HSP70), and cell division control protein 48 homolog A (CDC48) may play an important role in cabbage embryogenesis and were identified based on pathway enrichment and protein−protein interaction analyses. In addition, changes in the abundance of 9 representative DEPs were correlated with their corresponding mRNA levels using qRT-PCR. Carbohydrate metabolism supplies the energy needed for the rapid growth that occurs during embryo development, and the folding of synthesized proteins or the refolding of damaged and unstable proteins occur, which may due to the stress induced by in vitro culture. Conclusions A set of putative proteins presumably specific for microspore embryogenesis induced by high temperature treatment were identified. In isolated microspore culture of cabbage, we present the first exposition of non-embryo and the embryo (induced by 32 °C heat shock treatment 24h) changes in the expression of specific proteins.

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