iTRAQ-based quantitative proteomic analysis of cultivated Pseudostellaria heterophylla and its wild-type

Hua, Yujiao; Wang, Shengnan; Liu, Zixiu; Liu, Xunhong; Zou, Lisi; Gu, Wei; Hou, Yunlong; Ma, Yang; Luo, Yi‐Bo; Liu, Juanxiu

doi:10.1016/j.jprot.2016.02.027

Cited by 37 publications

(17 citation statements)

References 56 publications

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“…In order to depurate the protein extraction and determine the final protein concentration, the 2D Clean‐up Kit (GE Healthcare, UK) and 2D Quant Kit (GE Healthcare, London, UK) were used sequentially, following the manufacturer's instructions. Proteins were digested according to the FASP method . Protein samples (100 μg) were reduced using 2 μL of reducing agent and incubated at 60 °C for 1 h. Next, added 1 μL of cysteine‐blocking reagent at room temperature for 10 min using iTRAQ Reagent Multiplex Buffer Kit (AB Sciex, USA), and then transferred for ultrafiltration.…”

Section: Methodsmentioning

confidence: 99%

Analysis of Differentially Expressed Proteins Involved in Autoimmune Cirrhosis and Normal Serum by iTRAQ Proteomics

Zheng

Bao

et al. 2018

Proteomics Clinical Apps

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Section: Methodsmentioning

confidence: 99%

Analysis of Differentially Expressed Proteins Involved in Autoimmune Cirrhosis and Normal Serum by iTRAQ Proteomics

Zheng

Bao

et al. 2018

Proteomics Clinical Apps

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“…Elution was monitored by measuring the absorbance at 214 nm/280 nm, and fractions were collected every 1 min (Wang et al, ). The eluted peptides were pooled into 24 fractions, which were vacuum‐dried and acidized in 50% TFA for further nanoLC‐MS/MS analysis (Hua et al, ).…”

Section: Methodsmentioning

confidence: 99%

Quantitative proteomic analysis of intracerebral hemorrhage in rats with a focus on brain energy metabolism

et al. 2018

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IntroductionIntracerebral hemorrhage (ICH) is a lethal cerebrovascular disorder with a high mortality and morbidity. The pathophysiological mechanisms underlying ICH‐induced secondary injury remain unclear.MethodsTo examine one of the gaps in the knowledge about ICH pathological mechanisms, isobaric tag for relative and absolute quantification (iTRAQ)‐based liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) was used in collagenase‐induced ICH rats on the 2nd day.ResultsA total of 6,456 proteins were identified with a 1% false discovery rate (FDR). Of these proteins, 126 and 75 differentially expressed proteins (DEPs) were substantially increased and decreased, respectively. Based on Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and STRING analyses, the protein changes in cerebral hemorrhage were comprehensively evaluated, and the energy metabolism in ICH was anchored. The core position of the nitrogen metabolism pathway in brain metabolism in ICH was found for the first time. Carbonic anhydrase 1 (Ca1), carbonic anhydrase 2 (Ca2), and glutamine synthetase (Glul) participated in this pathway. We constructed the protein–protein interaction (PPI) networks for the energy metabolism of ICH, including the Atp6v1a‐Atp6v0c‐Atp6v0d1‐Ppa2‐Atp6ap2 network.ConclusionsIt seems that dysregulation of energy metabolism, especially nitrogen metabolism, may be a major cause in secondary ICH injury. This information provides novel insights into secondary events following ICH.

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“…Then the acquired sample was re-suspended with 20μl of 2% carbinol, 0.1% formic acid (12,000r, 10min, RT), and 10μl of supernatant was loaded onto EASY-Spray column (12cm×75μm, C18, 3μm), and a loading pump ow rate was 350 nl/min for 15 min and a separation ow rate was 350 nl/min. The separation gradient was as follows: 0 min, 4% mobile phase B (100% acetonitrile, 0.1% formic acid); 5 min, 15% mobile phase B; 40 min, 25% mobile phase B; 65 min, 35% mobile phase B; 70 min, 95% mobile phase B; 82 min, 95% mobile phase B; 85 min, 4% mobile phase B; 90 min, 4% mobile phase B, as previously described [15][16][17].…”

Section: Separation Of Peptides and Lc-ms/ms Analysismentioning

confidence: 99%

Proteome Analysis of Testicular Tissue in Varicocele Rats Using iTRAQ Labeling Technology

feng

Zhao

rong

et al. 2020

Preprint

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Purpose Varicocele (VC) is considered as the main cause of male infertility, clear and definite molecular markers of varicocele disease are helpful for early prevention and timely treatment. This study was in order to examine the effect of varicocele on protein expression in testicular tissue.Methods The testicular tissue samples of normal rats and varicocele rats were used for proteomic analysis and functional bioinformatics analysis. The proteins with a fold change of > 1.2 or < 1.2, and with a P-value < 0.05 were used to identify up- and downregulation proteins between the varicocele and control rats, and two target proteins were selected and verified with western blot.Results It was found that seminiferous epithelium was disordered and spermatogenic cells were injured seriously in varicocele group. A total of 65 differentially expressed proteins were identified compared with normal group by liquid chromatography tandem mass spectrometry (LC-MS/MS) and isobaric tags for relative and absolute quantitation (iTRAQ) analysis, including 31 up-regulated proteins and 34 down-regulated proteins, respectively. Functions of those proteins were mainly related to the following processes: signal transduction, protein cycle, transmembrane transport, protein transport, vesicular mediated transport, cell division, and membrane tissue. Two down regulated proteins of ATPase and Cu2+-transporting alpha (ATP7A) and Calcium and integrin-binding protein 1 (CIB1) in varicocele rats were selected and confirmed.Conclusion Protein spectrum of testicular tissue in varicocele rats was different from that in normal rats. Low expressions of ATP7A and CIB1 may be affect testicular spermatogenic function and can be used as a potential biomarker for testicular tissue to maintain normal spermatogenic function.

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iTRAQ-based quantitative proteomic analysis of cultivated Pseudostellaria heterophylla and its wild-type

Cited by 37 publications

References 56 publications

Analysis of Differentially Expressed Proteins Involved in Autoimmune Cirrhosis and Normal Serum by iTRAQ Proteomics

Analysis of Differentially Expressed Proteins Involved in Autoimmune Cirrhosis and Normal Serum by iTRAQ Proteomics

Quantitative proteomic analysis of intracerebral hemorrhage in rats with a focus on brain energy metabolism

Proteome Analysis of Testicular Tissue in Varicocele Rats Using iTRAQ Labeling Technology

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