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Proteomic analysis of human periodontal ligament cells under hypoxia 缺氧条件下人牙周膜细胞的蛋白质组学分析
IF 2 3区 生物学
Proteome Science Pub Date : 2019-08-31 DOI: 10.1186/s12953-019-0151-2
Qiwen Li, T. Luo, Wenxin Lu, X. Yi, Zhihe Zhao, Jun Liu
{"title":"Proteomic analysis of human periodontal ligament cells under hypoxia","authors":"Qiwen Li, T. Luo, Wenxin Lu, X. Yi, Zhihe Zhao, Jun Liu","doi":"10.1186/s12953-019-0151-2","DOIUrl":"https://doi.org/10.1186/s12953-019-0151-2","url":null,"abstract":"","PeriodicalId":20857,"journal":{"name":"Proteome Science","volume":"17 1","pages":""},"PeriodicalIF":2.0,"publicationDate":"2019-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s12953-019-0151-2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42152340","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 15
Comparison of enzymatic activities and proteomic profiles of Butyrivibrio fibrisolvens grown on different carbon sources. 在不同碳源上生长的溶纤维丁酸弧菌的酶活性和蛋白质组学比较。
IF 2 3区 生物学
Proteome Science Pub Date : 2019-06-01 eCollection Date: 2019-01-01 DOI: 10.1186/s12953-019-0150-3
Hana Sechovcová, Lucie Kulhavá, Kateřina Fliegerová, Mária Trundová, Daniel Morais, Jakub Mrázek, Jan Kopečný
{"title":"Comparison of enzymatic activities and proteomic profiles of <i>Butyrivibrio fibrisolvens</i> grown on different carbon sources.","authors":"Hana Sechovcová,&nbsp;Lucie Kulhavá,&nbsp;Kateřina Fliegerová,&nbsp;Mária Trundová,&nbsp;Daniel Morais,&nbsp;Jakub Mrázek,&nbsp;Jan Kopečný","doi":"10.1186/s12953-019-0150-3","DOIUrl":"https://doi.org/10.1186/s12953-019-0150-3","url":null,"abstract":"<p><strong>Background: </strong>The rumen microbiota is one of the most complex consortia of anaerobes, involving archaea, bacteria, protozoa, fungi and phages. They are very effective at utilizing plant polysaccharides, especially cellulose and hemicelluloses. The most important hemicellulose decomposers are clustered with the genus <i>Butyrivibrio</i>. As the related species differ in their range of hydrolytic activities and substrate preferences, <i>Butyrivibrio fibrisolvens</i> was selected as one of the most effective isolates and thus suitable for proteomic studies on substrate comparisons in the extracellular fraction. The <i>B. fibrisolvens</i> genome is the biggest in the butyrivibria cluster and is focused on \"environmental information processing\" and \"carbohydrate metabolism\".</p><p><strong>Methods: </strong>The study of the effect of carbon source on <i>B. fibrisolvens</i> 3071 was based on cultures grown on four substrates: xylose, glucose, xylan, xylan with 25% glucose. The enzymatic activities were studied by spectrophotometric and zymogram methods. Proteomic study was based on genomics, 2D electrophoresis and nLC/MS (Bruker Daltonics) analysis.</p><p><strong>Results: </strong>Extracellular β-endoxylanase as well as xylan β-xylosidase activities were induced with xylan. The presence of the xylan polymer induced hemicellulolytic enzymes and increased the protein fraction in the interval from 40 to 80 kDa. 2D electrophoresis with nLC/MS analysis of extracellular <i>B. fibrisolvens</i> 3071 proteins found 14 diverse proteins with significantly different expression on the tested substrates.</p><p><strong>Conclusion: </strong>The comparison of four carbon sources resulted in the main significant changes in <i>B. fibrisolvens</i> proteome occurring outside the fibrolytic cluster of proteins. The affected proteins mainly belonged to the glycolysis and protein synthesis cluster.</p>","PeriodicalId":20857,"journal":{"name":"Proteome Science","volume":" ","pages":"2"},"PeriodicalIF":2.0,"publicationDate":"2019-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s12953-019-0150-3","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40448024","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 7
Hepatic protein Carbonylation profiles induced by lipid accumulation and oxidative stress for investigating cellular response to non-alcoholic fatty liver disease in vitro. 脂质积聚和氧化应激诱导的肝蛋白羰基化谱用于研究细胞对非酒精性脂肪性肝病的体外反应。
IF 2.1 3区 生物学
Proteome Science Pub Date : 2019-03-27 eCollection Date: 2019-01-01 DOI: 10.1186/s12953-019-0149-9
Peerut Chienwichai, Onrapak Reamtong, Usa Boonyuen, Trairak Pisitkun, Poorichaya Somparn, Prapin Tharnpoophasiam, Suwalee Worakhunpiset, Supachai Topanurak
{"title":"Hepatic protein Carbonylation profiles induced by lipid accumulation and oxidative stress for investigating cellular response to non-alcoholic fatty liver disease in vitro.","authors":"Peerut Chienwichai, Onrapak Reamtong, Usa Boonyuen, Trairak Pisitkun, Poorichaya Somparn, Prapin Tharnpoophasiam, Suwalee Worakhunpiset, Supachai Topanurak","doi":"10.1186/s12953-019-0149-9","DOIUrl":"10.1186/s12953-019-0149-9","url":null,"abstract":"<p><strong>Background: </strong>Non-alcoholic fatty liver disease (NAFLD) is caused by excessive accumulation of fat within the liver, leading to further severe conditions such as non-alcoholic steatohepatitis (NASH). Progression of healthy liver to steatosis and NASH is not yet fully understood in terms of process and response. Hepatic oxidative stress is believed to be one of the factors driving steatosis to NASH. Oxidative protein modification is the major cause of protein functional impairment in which alteration of key hepatic enzymes is likely to be a crucial factor for NAFLD biology. In the present study, we aimed to discover carbonylated protein profiles involving in NAFLD biology in vitro.</p><p><strong>Methods: </strong>Hepatocyte cell line was used to induce steatosis with fatty acids (FA) in the presence and absence of menadione (oxidative stress inducer). Two-dimensional gel electrophoresis-based proteomics and dinitrophenyl hydrazine derivatization technique were used to identify carbonylated proteins. Sequentially, in order to view changes in protein carbonylation pathway, enrichment using Funrich algorithm was performed. The selected carbonylated proteins were validated with western blot and carbonylated sites were further identified by high-resolution LC-MS/MS.</p><p><strong>Results: </strong>Proteomic results and pathway analysis revealed that carbonylated proteins are involved in NASH pathogenesis pathways in which most of them play important roles in energy metabolisms. Particularly, carbonylation level of ATP synthase subunit α (ATP5A), a key protein in cellular respiration, was reduced after FA and FA with oxidative stress treatment, whereas its expression was not altered. Carbonylated sites on this protein were identified and it was revealed that these sites are located in nucleotide binding region. Modification of these sites may, therefore, disturb ATP5A activity. As a consequence, the lower carbonylation level on ATP5A after FA treatment solely or with oxidative stress can increase ATP production.</p><p><strong>Conclusions: </strong>The reduction in carbonylated level of ATP5A might occur to generate more energy in response to pathological conditions, in our case, fat accumulation and oxidative stress in hepatocytes. This would imply the association between protein carbonylation and molecular response to development of steatosis and NASH.</p>","PeriodicalId":20857,"journal":{"name":"Proteome Science","volume":"17 ","pages":"1"},"PeriodicalIF":2.1,"publicationDate":"2019-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6438040/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37132632","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Proteome modifications on tomato under extreme high light induced-stress. 极端强光诱导胁迫下番茄的蛋白质组变化。
IF 2 3区 生物学
Proteome Science Pub Date : 2018-11-24 eCollection Date: 2018-01-01 DOI: 10.1186/s12953-018-0148-2
Débora Parrine, Bo-Sen Wu, Bilal Muhammad, Keith Rivera, Darryl Pappin, Xin Zhao, Mark Lefsrud
{"title":"Proteome modifications on tomato under extreme high light induced-stress.","authors":"Débora Parrine, Bo-Sen Wu, Bilal Muhammad, Keith Rivera, Darryl Pappin, Xin Zhao, Mark Lefsrud","doi":"10.1186/s12953-018-0148-2","DOIUrl":"10.1186/s12953-018-0148-2","url":null,"abstract":"<p><strong>Background: </strong>Abiotic stress reduces photosynthetic yield and plant growth, negatively impacting global crop production and is a major constraint faced by agriculture. However, the knowledge on the impact on plants under extremely high irradiance is limited. We present the first in-depth proteomics analysis of plants treated with a method developed by our research group to generate a light gradient using an extremely intense light.</p><p><strong>Methods: </strong>The method consists of utilizing light emitting diodes (LED) to create a single spot at 24,000 μmol m<sup>- 2</sup> s<sup>- 1</sup> irradiance, generating three light stress levels. A light map and temperature profile were obtained during the light experiment. The proteins expressed in the treated tomato (<i>Solanum lycopersicum</i>, Heinz H1706) leaves were harvested 10 days after the treatment, allowing for the detection of proteins involved in a long-term recovery. A multiplex labeled proteomics method (iTRAQ) was analyzed by LC-MS/MS.</p><p><strong>Results: </strong>A total of 3994 proteins were identified at 1% false discovery rate and matched additional quality filters. Hierarchical clustering analysis resulted in four types of patterns related to the protein expression, with one being directly linked to the increased LED irradiation. A total of 37 proteins were found unique to the least damaged leaf zone, while the medium damaged zone had 372 proteins, and the severely damaged presented unique 1003 proteins. Oxygen evolving complex and PSII complex proteins (PsbH, PsbS, PsbR and Psb28) were found to be abundant in the most damaged leaf zone. This leaf zone presented a protein involved in the salicylic acid response, while it was not abundant in the other leaf zones. The mRNA level of PsbR was significantly lower (1-fold) compared the control in the most damaged zone of the leaf, while Psb28 and PsbH were lower (1-fold) in the less damaged leaf zones. PsbS mRNA abundance in all leaf zones tested presented no statistically significant change from the control.</p><p><strong>Conclusions: </strong>We present the first characterization of the proteome changes caused by an extreme level of high-light intensity (24,000 μmol m<sup>- 2</sup> s<sup>- 1</sup>). The proteomics results show the presence of specific defense responses to each level of light intensity, with a possible involvement of proteins PsbH, Psb28, PsbR, and PsbS.</p>","PeriodicalId":20857,"journal":{"name":"Proteome Science","volume":"16 ","pages":"20"},"PeriodicalIF":2.0,"publicationDate":"2018-11-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6260845/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36768405","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Key factors identified by proteomic analysis in maize (Zea mays L.) seedlings' response to long-term exposure to different phosphate levels. 玉米(Zea mays L.)幼苗对不同磷水平长期暴露响应的蛋白质组学分析鉴定关键因素。
IF 2 3区 生物学
Proteome Science Pub Date : 2018-11-20 eCollection Date: 2018-01-01 DOI: 10.1186/s12953-018-0147-3
Yanling Sun, Chunhua Mu, Xia Liu
{"title":"Key factors identified by proteomic analysis in maize (<i>Zea mays</i> L.) seedlings' response to long-term exposure to different phosphate levels.","authors":"Yanling Sun,&nbsp;Chunhua Mu,&nbsp;Xia Liu","doi":"10.1186/s12953-018-0147-3","DOIUrl":"https://doi.org/10.1186/s12953-018-0147-3","url":null,"abstract":"<p><strong>Background: </strong>Maize seedlings are constantly exposed to inorganic phosphate (Pi)-limited environments. To understand how maize cope with low Pi (LP) and high Pi (HP) conditions, physiological and global proteomic analysis of QXN233 genotype were performed under the long-term Pi starvation and supplementation.</p><p><strong>Methods: </strong>We investigated the physiological response of QXN233 genotype to LP and HP conditions and detected the changes in ion fluxes by non-invasive micro-test technology and gene expression by quantitative real-time polymerase chain reaction. QXN233 was further assessed using vermiculite assay, and then proteins were isolated and identified by nano-liquid chromatography-mass spectrometry.</p><p><strong>Results: </strong>A negative relationship was observed between Na<sup>+</sup> and Pi, and Na<sup>+</sup> efflux was enhanced under HP condition. Furthermore, a total of 681 and 1374 were identified in the leaves and roots, respectively, which were mostly involved in metabolism, ion transport, and stress response. Importantly, several key Pi transporters were identified for breeding potential. Several ion transporters demonstrated an elaborate interplay between Pi and other ions, together contributing to the growth of QXN233 seedlings.</p><p><strong>Conclusion: </strong>The results from this study provide insights into the response of maize seedlings to long-term Pi exposure.</p>","PeriodicalId":20857,"journal":{"name":"Proteome Science","volume":"16 ","pages":"19"},"PeriodicalIF":2.0,"publicationDate":"2018-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s12953-018-0147-3","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36721576","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 9
Calcitonin gene-related peptide promotes proliferation and inhibits apoptosis in endothelial progenitor cells via inhibiting MAPK signaling. 降钙素基因相关肽通过抑制MAPK信号传导促进内皮祖细胞增殖和抑制细胞凋亡。
IF 2 3区 生物学
Proteome Science Pub Date : 2018-11-14 eCollection Date: 2018-01-01 DOI: 10.1186/s12953-018-0146-4
Jianqun Wu, Song Liu, Zhao Wang, Shenghui Ma, Huan Meng, Jijie Hu
{"title":"Calcitonin gene-related peptide promotes proliferation and inhibits apoptosis in endothelial progenitor cells via inhibiting MAPK signaling.","authors":"Jianqun Wu,&nbsp;Song Liu,&nbsp;Zhao Wang,&nbsp;Shenghui Ma,&nbsp;Huan Meng,&nbsp;Jijie Hu","doi":"10.1186/s12953-018-0146-4","DOIUrl":"https://doi.org/10.1186/s12953-018-0146-4","url":null,"abstract":"<p><strong>Background: </strong>Calcitonin gene-related peptide (CGRP) contributes to bone formation by stimulating bone marrow stromal cell (BMSC) proliferation and differentiation. However, the proliferative and apoptotic effects of CGRP on bone marrow-derived endothelial progenitor cells (EPCs) have not been investigated.</p><p><strong>Methods: </strong>We tested the effects of CGRP on EPC proliferation and apoptosis by Cell Counting Kit-8, flow cytometry, and studied the effects of CGRP on the expression of proliferation- and apoptosis-related markers in EPCs and the underlying mitogen-activated protein kinase (MAPK) signalling pathway by quantitative polymerase chain reaction and western blotting.</p><p><strong>Results: </strong>We detected EPC markers (CD34, CD133 and VEGFR-2) in 7-day cultures and found that CGRP (10<sup>- 10</sup>-10<sup>- 12</sup> M) promoted the proliferation of cultured EPCs, with a peak increase of 30% at 10<sup>- 10</sup> M CGRP. CGRP also upregulated the expression of proliferation-associated genes, including cyclin D1 and cyclin E, and increased the percentages of G2/M-phase and S-phase cells after incubation 72 h. CGRP inhibited serum deprivation (SD)-induced apoptosis in EPCs after 24 and 48 h and downregulated the expression of apoptosis-related genes, including caspase-3, caspase-8, caspase-9 and Bax. Phosphorylated (p-)ERK1/2, p-p38 and p-JNK protein levels in EPCs treated with CGRP were significantly lower than those in untreated EPCs. Pre-treatment with the calcitonin receptor-like receptor (CRLR) antagonist CGRP8-37 or a MAPK pathway inhibitor (PD98059, SB203580 or SP600125) completely or partially reversed the pro-proliferative and anti-apoptotic effects and the reduced p-ERK1/2, p-p38 and p-JNK expression induced by CGRP.</p><p><strong>Conclusion: </strong>Our results show that CGRP exerts pro-proliferative and anti-apoptotic effects on EPCs and may act by inhibiting MAPK pathways.</p>","PeriodicalId":20857,"journal":{"name":"Proteome Science","volume":"16 ","pages":"18"},"PeriodicalIF":2.0,"publicationDate":"2018-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s12953-018-0146-4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36763011","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 16
Comparative proteomic analysis of Ulva prolifera response to high temperature stress. 增生Ulva对高温胁迫反应的比较蛋白质组学分析。
IF 2 3区 生物学
Proteome Science Pub Date : 2018-10-27 eCollection Date: 2018-01-01 DOI: 10.1186/s12953-018-0145-5
Meihua Fan, Xue Sun, Zhi Liao, Jianxin Wang, Yahe Li, Nianjun Xu
{"title":"Comparative proteomic analysis of <i>Ulva prolifera</i> response to high temperature stress.","authors":"Meihua Fan,&nbsp;Xue Sun,&nbsp;Zhi Liao,&nbsp;Jianxin Wang,&nbsp;Yahe Li,&nbsp;Nianjun Xu","doi":"10.1186/s12953-018-0145-5","DOIUrl":"https://doi.org/10.1186/s12953-018-0145-5","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;&lt;i&gt;Ulva prolifera&lt;/i&gt; belongs to green macroalgae and is the dominant species of green tide. It is distributed worldwide and is therefore subject to high-temperature stress during the growth process. However, the adaptation mechanisms of the response of &lt;i&gt;U. prolifera&lt;/i&gt; to high temperatures have not been clearly investigated yet.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;In this study, isobaric tags for relative and absolute quantitation (iTRAQ) labelling was applied in combination with the liquid chromatography-tandem mass spectrometry (LC-MS/MS) to conduct comparative proteomic analysis of the response of &lt;i&gt;U. prolifera&lt;/i&gt; to high-temperature stress and to elucidate the involvement of this response in adaptation mechanisms. Differentially expressed proteins (DEPs) of &lt;i&gt;U. prolifera&lt;/i&gt; under high temperature (denote UpHT) compared with the control (UpC) were identified. Bioinformatic analyses including GO analysis, pathway analysis, and pathway enrichment analysis was performed to analyse the key metabolic pathways that underlie the thermal tolerance mechanism through protein networks. Quantitative real-time PCR and western blot were performed to validate selected proteins.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;In the present study, 1223 DEPs were identified under high temperature compared with the control, which included 790 up-regulated and 433 down-regulated proteins. The high-temperature stimulus mainly induced the expression of glutathione S-transferase, heat shock protein, ascorbate peroxidase, manganese superoxide dismutase, ubiquitin-related protein, lhcSR, rubisco activase, serine/threonine protein kinase 2, adenylate kinase, Ca&lt;sup&gt;2+&lt;/sup&gt;-dependent protein kinase (CDPK), disease resistance protein EDS1, metacaspase type II, NDPK2a, 26S proteasome regulatory subunit, ubiquinone oxidoreductase, ATP synthase subunit, SnRK2s, and cytochrome P450. The down-regulated proteins were photosynthesis-related proteins, glutathione reductase, catalase-peroxidase, thioredoxin, thioredoxin peroxidase, PP2C, and carbon fixation-related proteins. Furthermore, biological index analysis indicated that protein content and SOD activity decreased; the value of Fv/Fm dropped to the lowest point after culture for 96 h. However, APX activity and MDA content increased under high temperature.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusion: &lt;/strong&gt;The present study implied an increase in proteins that were associated with the stress response, oxidative phosphorylation, the cytokinin signal transduction pathway, the abscisic acid signal transduction pathway, and the glutathione metabolism pathway. Proteins that were associated with photosynthesis, carbon fixation in photosynthesis organisms, and the photosynthesis antenna protein pathway were decreased. These pathways played a pivotal role in high temperature regulation. These novel proteins provide a good starting point for further research into their functions using genetic or other approaches. These findin","PeriodicalId":20857,"journal":{"name":"Proteome Science","volume":"16 ","pages":"17"},"PeriodicalIF":2.0,"publicationDate":"2018-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s12953-018-0145-5","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36639097","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 19
Global phosphoproteomic analysis identifies SRMS-regulated secondary signaling intermediates. 全球磷酸化蛋白质组学分析确定了srms调控的二级信号中间体。
IF 2 3区 生物学
Proteome Science Pub Date : 2018-08-18 eCollection Date: 2018-01-01 DOI: 10.1186/s12953-018-0143-7
Raghuveera Kumar Goel, Mona Meyer, Marta Paczkowska, Jüri Reimand, Frederick Vizeacoumar, Franco Vizeacoumar, TuKiet T Lam, Kiven Erique Lukong
{"title":"Global phosphoproteomic analysis identifies SRMS-regulated secondary signaling intermediates.","authors":"Raghuveera Kumar Goel,&nbsp;Mona Meyer,&nbsp;Marta Paczkowska,&nbsp;Jüri Reimand,&nbsp;Frederick Vizeacoumar,&nbsp;Franco Vizeacoumar,&nbsp;TuKiet T Lam,&nbsp;Kiven Erique Lukong","doi":"10.1186/s12953-018-0143-7","DOIUrl":"https://doi.org/10.1186/s12953-018-0143-7","url":null,"abstract":"<p><strong>Background: </strong>The non-receptor tyrosine kinase, SRMS (Src-related kinase lacking C-terminal regulatory tyrosine and N-terminal myristoylation sites) is a member of the BRK family kinases (BFKs) which represents an evolutionarily conserved relative of the Src family kinases (SFKs). Tyrosine kinases are known to regulate a number of cellular processes and pathways via phosphorylating substrate proteins directly and/or by partaking in signaling cross-talks leading to the indirect modulation of various signaling intermediates. In a previous study, we profiled the tyrosine-phosphoproteome of SRMS and identified multiple candidate substrates of the kinase. The broader cellular signaling intermediates of SRMS are unknown.</p><p><strong>Methods: </strong>In order to uncover the broader SRMS-regulated phosphoproteome and identify the SRMS-regulated indirect signaling intermediates, we performed label-free global phosphoproteomics analysis on cells expressing wild-type SRMS. Using computational database searching and bioinformatics analyses we characterized the dataset.</p><p><strong>Results: </strong>Our analyses identified 60 hyperphosphorylated (phosphoserine/phosphothreonine) proteins mapped from 140 hyperphosphorylated peptides. Bioinfomatics analyses identified a number of significantly enriched biological and cellular processes among which DNA repair pathways were found to be upregulated while apoptotic pathways were found to be downregulated. Analyses of motifs derived from the upregulated phosphosites identified Casein kinase 2 alpha (CK2α) as one of the major potential kinases contributing to the SRMS-dependent indirect regulation of signaling intermediates.</p><p><strong>Conclusions: </strong>Overall, our phosphoproteomics analyses identified serine/threonine phosphorylation dynamics as important secondary events of the SRMS-regulated phosphoproteome with implications in the regulation of cellular and biological processes.</p>","PeriodicalId":20857,"journal":{"name":"Proteome Science","volume":"16 ","pages":"16"},"PeriodicalIF":2.0,"publicationDate":"2018-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s12953-018-0143-7","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36425320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 11
Label-free quantitative mass spectrometry analysis of differential protein expression in the developing cochlear sensory epithelium. 发育中的耳蜗感觉上皮中差异蛋白表达的无标记定量质谱分析。
IF 2 3区 生物学
Proteome Science Pub Date : 2018-08-07 eCollection Date: 2018-01-01 DOI: 10.1186/s12953-018-0144-6
Lancia N F Darville, Bernd H A Sokolowski
{"title":"Label-free quantitative mass spectrometry analysis of differential protein expression in the developing cochlear sensory epithelium.","authors":"Lancia N F Darville,&nbsp;Bernd H A Sokolowski","doi":"10.1186/s12953-018-0144-6","DOIUrl":"https://doi.org/10.1186/s12953-018-0144-6","url":null,"abstract":"<p><strong>Background: </strong>The sensory epithelium of the inner ear converts the mechanical energy of sound to electro-chemical energy recognized by the central nervous system. This process is mediated by receptor cells known as hair cells that express proteins in a timely fashion with the onset of hearing.</p><p><strong>Methods: </strong>The proteomes of 3, 14, and 30 day-old mice cochlear sensory epithelia were revealed, using label-free quantitative mass spectrometry (LTQ-Orbitrap). Statistical analysis using a one-way ANOVA followed by Bonferroni's <i>post-hoc</i> test was used to show significant differences in protein expression. Ingenuity Pathway Analysis was used to observe networks of differentially expressed proteins, their biological processes, and associated diseases, while Cytoscape software was used to determine putative interactions with select biomarker proteins. These candidate biomarkers were further verified using Western blotting, while coimmunoprecipitation was used to verify putative partners determined using bioinformatics.</p><p><strong>Results: </strong>We show that a comparison across all three proteomes shows that there are 447 differentially expressed proteins, with 387 differentially expressed between postnatal day 3 and 30. Ingenuity Pathway Analysis revealed ~ 62% of postnatal day 3 downregulated proteins are involved in neurological diseases. Several proteins are expressed exclusively on P3, including Parvin α, Drebrin1 (Drb1), Secreted protein acidic and cysteine rich (SPARC), Transmembrane emp24 domain-containing protein 10 (Tmed10). Coimmunoprecipitations showed that Parvin and SPARC interact with integrin-linked protein kinase and the large conductance calcium-activated potassium channel, respectively.</p><p><strong>Conclusions: </strong>Quantitative mass spectrometry revealed the identification of numerous differentially regulated proteins over three days of postnatal development. These data provide insights into functional pathways regulating normal sensory and supporting cell development in the cochlea that include potential biomarkers. Interacting partners of two of these markers suggest the importance of these complexes in regulating cellular structure and synapse development.</p>","PeriodicalId":20857,"journal":{"name":"Proteome Science","volume":"16 ","pages":"15"},"PeriodicalIF":2.0,"publicationDate":"2018-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s12953-018-0144-6","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36410411","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 5
Comparative iTRAQ proteomics revealed proteins associated with horn development in yak. 比较iTRAQ蛋白质组学揭示了牦牛角发育的相关蛋白。
IF 2 3区 生物学
Proteome Science Pub Date : 2018-07-24 eCollection Date: 2018-01-01 DOI: 10.1186/s12953-018-0141-9
Mingna Li, Xiaoyun Wu, Xian Guo, Pengjia Bao, Xuezhi Ding, Min Chu, Chunnian Liang, Ping Yan
{"title":"Comparative iTRAQ proteomics revealed proteins associated with horn development in yak.","authors":"Mingna Li,&nbsp;Xiaoyun Wu,&nbsp;Xian Guo,&nbsp;Pengjia Bao,&nbsp;Xuezhi Ding,&nbsp;Min Chu,&nbsp;Chunnian Liang,&nbsp;Ping Yan","doi":"10.1186/s12953-018-0141-9","DOIUrl":"https://doi.org/10.1186/s12953-018-0141-9","url":null,"abstract":"<p><strong>Background: </strong>The practice of dehorning yak raises animal safety concerns, which have been addressed by selective breeding to obtain genetically hornless yak. The POLLED locus in yak has been studied extensively; however, little is known regarding the proteins that regulate horn bud development.</p><p><strong>Methods: </strong>A differential proteomic analysis was performed to compare the skin from the horn bud region of polled yak fetuses and the horn bud tissue of horned yak fetuses using isobaric tags for relative and absolute quantitation (iTRAQ) technology coupled with 2D LC-MS/MS.</p><p><strong>Results: </strong>One hundred differentially abundant proteins (DAPs) were identified. Of these, 29 were up-regulated and 71 were down-regulated in skin from the horn bud region of polled fetuses when compared to the horn bud tissue of horned fetuses. Bioinformatics analyses showed that the up-regulated DAPs were mainly associated with metabolic activities, while the down-regulated DAPs were significantly enriched in cell adhesion and cell movement activities.</p><p><strong>Conclusions: </strong>We concluded that some important proteins were associated with cell adhesion, cell motility, keratinocyte differentiation, cytoskeleton organization, osteoblast differentiation, and fatty acid metabolism during horn bud development. These results advance our understanding of the molecular mechanisms underlying horn development.</p>","PeriodicalId":20857,"journal":{"name":"Proteome Science","volume":"16 ","pages":"14"},"PeriodicalIF":2.0,"publicationDate":"2018-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s12953-018-0141-9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36357905","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 13
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