Zhiwei Qiao, Takashi Tajima, F. Kito, Y. Arai, A. Kawai, T. Kondo
Myxoid liposarcoma (MLS) is a rare mesenchymal malignancy with unique extrapulmonary metastatic potential. Although MLS has been associated with specific chromosomal translocations, the factors and pathways regulating metastasis in MLS remain unknown. To identify the molecular mechanisms underlying MLS metastasis, we compared global gene expression profiles of primary tumor tissues from MLS patients with different metastatic statuses using DNA microarray analysis. In total, 393 genes were differentially expressed between the tumors from four patients with metastasis and those from 11 patients without metastasis. Differentially expressed genes were functionally annotated based on Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Supervised classification based on the 393 genes clearly discriminated samples according to metastatic status. The pathways responsible for metastasis included “focal adhesion,” “pathways in cancer,” “ECM-receptor interaction,” and “tight junction.” The differential expression of alpha-synuclein was confirmed at the protein level; the protein was downregulated in metastatic MLS. Meta-analysis revealed that MLS could be discriminated from the other sarcomas based on the expression of the metastasis-associated genes. The metastasis-associated genes identified in this study are worthwhile further investigation to further our understanding of MLS and are expected to lead to novel clinical applications for MLS.
{"title":"Metastasis-associated gene signature in primary myxoid liposarcoma identified through a gene expression study","authors":"Zhiwei Qiao, Takashi Tajima, F. Kito, Y. Arai, A. Kawai, T. Kondo","doi":"10.2198/JELECTROPH.61.9","DOIUrl":"https://doi.org/10.2198/JELECTROPH.61.9","url":null,"abstract":"Myxoid liposarcoma (MLS) is a rare mesenchymal malignancy with unique extrapulmonary metastatic potential. Although MLS has been associated with specific chromosomal translocations, the factors and pathways regulating metastasis in MLS remain unknown. To identify the molecular mechanisms underlying MLS metastasis, we compared global gene expression profiles of primary tumor tissues from MLS patients with different metastatic statuses using DNA microarray analysis. In total, 393 genes were differentially expressed between the tumors from four patients with metastasis and those from 11 patients without metastasis. Differentially expressed genes were functionally annotated based on Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Supervised classification based on the 393 genes clearly discriminated samples according to metastatic status. The pathways responsible for metastasis included “focal adhesion,” “pathways in cancer,” “ECM-receptor interaction,” and “tight junction.” The differential expression of alpha-synuclein was confirmed at the protein level; the protein was downregulated in metastatic MLS. Meta-analysis revealed that MLS could be discriminated from the other sarcomas based on the expression of the metastasis-associated genes. The metastasis-associated genes identified in this study are worthwhile further investigation to further our understanding of MLS and are expected to lead to novel clinical applications for MLS.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"164 1","pages":"9-15"},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77265571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zhiwei Qiao, F. Kito, Y. Takai, R. Oyama, T. Kondo
SUMMARY Sarcoma is a rare malignancy with an aggressive clinical course. Tyrosine kinase inhibitors (TKIs) have emerged as effective drugs in targeted therapy for malignancies; in particular, pazopanib was recently approved for the treatment of sarcoma. However, as only a limited proportion of patients exhibit favorable response to treatment with TKIs, predictive biomarkers of response to these drugs are urgently needed. In this study, we attempted to identify predictive biomarkers for response to TKIs in synovial sarcoma. We performed a magnetic bead–based assay (Bio-Plex) using synovial sarcoma cell culture supernatant and validated the results by ELISA and western blotting. Cellular protein and mRNA expression levels of candidate biomarkers were evaluated by western blotting and RT-PCR. Gene expression profiling of candidate biomarkers was conducted by meta-analysis of publicly available gene expression data from 149 patients with synovial sarcoma. We found that follistatin (FST) was significantly highly expressed in TKI-resistant cells. Moreover, cell proliferation was decreased following gene silencing of FST . Meta-analysis revealed that the mRNA expression of FST varied among the 149 patients with synovial sarcoma, and that 23 genes were co-expressed with FST ; these included genes encoding receptor tyrosine kinase-like orphan receptor 1, Sal-like protein 4, and signal transducer CD24. This study suggested that FST represents a candidate predictive biomarker for response to treatment with TKIs in synovial sarcoma. Secretomics is a promising approach for predictive biomarker exploration. The utility of FST as a predictive biomarker for response to treatment with TKIs in synovial sarcoma should be further validated using clinical samples.
{"title":"Secretomics identifies follistatin as a predictive biomarker for response to treatment with tyrosine kinase inhibitors in synovial sarcoma","authors":"Zhiwei Qiao, F. Kito, Y. Takai, R. Oyama, T. Kondo","doi":"10.2198/JELECTROPH.61.1","DOIUrl":"https://doi.org/10.2198/JELECTROPH.61.1","url":null,"abstract":"SUMMARY Sarcoma is a rare malignancy with an aggressive clinical course. Tyrosine kinase inhibitors (TKIs) have emerged as effective drugs in targeted therapy for malignancies; in particular, pazopanib was recently approved for the treatment of sarcoma. However, as only a limited proportion of patients exhibit favorable response to treatment with TKIs, predictive biomarkers of response to these drugs are urgently needed. In this study, we attempted to identify predictive biomarkers for response to TKIs in synovial sarcoma. We performed a magnetic bead–based assay (Bio-Plex) using synovial sarcoma cell culture supernatant and validated the results by ELISA and western blotting. Cellular protein and mRNA expression levels of candidate biomarkers were evaluated by western blotting and RT-PCR. Gene expression profiling of candidate biomarkers was conducted by meta-analysis of publicly available gene expression data from 149 patients with synovial sarcoma. We found that follistatin (FST) was significantly highly expressed in TKI-resistant cells. Moreover, cell proliferation was decreased following gene silencing of FST . Meta-analysis revealed that the mRNA expression of FST varied among the 149 patients with synovial sarcoma, and that 23 genes were co-expressed with FST ; these included genes encoding receptor tyrosine kinase-like orphan receptor 1, Sal-like protein 4, and signal transducer CD24. This study suggested that FST represents a candidate predictive biomarker for response to treatment with TKIs in synovial sarcoma. Secretomics is a promising approach for predictive biomarker exploration. The utility of FST as a predictive biomarker for response to treatment with TKIs in synovial sarcoma should be further validated using clinical samples.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"56 1","pages":"1-7"},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79826877","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Toshihiro Yoshimura, K. Kurogi, Ming-Cheh Liu, M. Suiko, Y. Sakakibara
S-nitrosylation, a post-translational modification of the thiol group of cysteine residues by nitric oxide (NO), has emerged as a new mode of signal transduction and regulation of protein function. It has recently been shown that S-nitrosylation may result in various protein dysfunctions. However, an improved S-nitrosylation analysis method is needed to achieve high sensitivity and quantitative accuracy. We hypothesized that an analysis method using fluorescence dye could detect S-nitrosylated proteins at a higher sensitivity than that of the conventional method. In this study, we developed a procedure for analyzing S-nitrosylated proteins using CyDye (the CyDye switch method). This CyDye switch method for detecting S-nitrosylated proteins was developed based on the biotin-switch method for analyzing S-nitrosylated proteins. We analyzed NO donor-induced S-nitrosylated proteins in a model protein and at the cellular level. We demonstrated that this CyDye switch method could detect specific S-nitrosylated proteins using SDS-PAGE and mass spectrometry. Our results indicate that the optimized CyDye switch method is suitable for the detection of the post-translational S-nitrosylation of proteins.
{"title":"A proteomic approach for the analysis of S-nitrosylated proteins using a fluorescence labeling technique","authors":"Toshihiro Yoshimura, K. Kurogi, Ming-Cheh Liu, M. Suiko, Y. Sakakibara","doi":"10.2198/JELECTROPH.60.5","DOIUrl":"https://doi.org/10.2198/JELECTROPH.60.5","url":null,"abstract":"S-nitrosylation, a post-translational modification of the thiol group of cysteine residues by nitric oxide (NO), has emerged as a new mode of signal transduction and regulation of protein function. It has recently been shown that S-nitrosylation may result in various protein dysfunctions. However, an improved S-nitrosylation analysis method is needed to achieve high sensitivity and quantitative accuracy. We hypothesized that an analysis method using fluorescence dye could detect S-nitrosylated proteins at a higher sensitivity than that of the conventional method. In this study, we developed a procedure for analyzing S-nitrosylated proteins using CyDye (the CyDye switch method). This CyDye switch method for detecting S-nitrosylated proteins was developed based on the biotin-switch method for analyzing S-nitrosylated proteins. We analyzed NO donor-induced S-nitrosylated proteins in a model protein and at the cellular level. We demonstrated that this CyDye switch method could detect specific S-nitrosylated proteins using SDS-PAGE and mass spectrometry. Our results indicate that the optimized CyDye switch method is suitable for the detection of the post-translational S-nitrosylation of proteins.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"36 1","pages":"5-14"},"PeriodicalIF":0.0,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73178843","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Protein post-translational modifications (PTMs) play crucial roles in regulation of protein function and cell signaling, and abnormalities in protein PTMs are both causes and consequences of disease. Mass spectrometry (MS) is widely used to analyze protein PTMs. In this study, we developed an original database, ModProt (Post-Translational Modification Map of Proteome), to integrate our laboratory data and literature information regarding PTM sites. To develop the ModProt database, we constructed a web-based laboratory information management system (LIMS). This system allows us to administer the ModProt database and to view PTM site maps and corresponding protein information including amino acid sequences, official gene symbols, UniProt accessions/IDs, chromosome number/ positions, and additional description. The ultimate goal of the ModProt database is to achieve PTM-based diagnosis and personalized medicine through detection of abnormal PTMs by comparing PTM site maps in healthy and disease states using the database.
蛋白质翻译后修饰(PTMs)在蛋白质功能和细胞信号传导调节中起着至关重要的作用，蛋白质PTMs的异常既是疾病的原因也是疾病的后果。质谱法(MS)广泛用于蛋白质PTMs的分析。在这项研究中，我们开发了一个原始数据库ModProt (Post-Translational Modification Map of Proteome)，来整合我们关于PTM位点的实验室数据和文献信息。为了开发ModProt数据库，我们构建了一个基于web的实验室信息管理系统(LIMS)。该系统允许我们管理ModProt数据库，并查看PTM位点图和相应的蛋白质信息，包括氨基酸序列、官方基因符号、UniProt加入/ id、染色体数目/位置和其他描述。ModProt数据库的最终目标是通过比较数据库中健康状态和疾病状态下的PTM位点图，发现异常的PTM，从而实现基于PTM的诊断和个性化医疗。
{"title":"ModProt: a database for integrating laboratory and literature data about protein post-translational modifications","authors":"Y. Kimura, T. Toda, H. Hirano","doi":"10.2198/JELECTROPH.60.1","DOIUrl":"https://doi.org/10.2198/JELECTROPH.60.1","url":null,"abstract":"Protein post-translational modifications (PTMs) play crucial roles in regulation of protein function and cell signaling, and abnormalities in protein PTMs are both causes and consequences of disease. Mass spectrometry (MS) is widely used to analyze protein PTMs. In this study, we developed an original database, ModProt (Post-Translational Modification Map of Proteome), to integrate our laboratory data and literature information regarding PTM sites. To develop the ModProt database, we constructed a web-based laboratory information management system (LIMS). This system allows us to administer the ModProt database and to view PTM site maps and corresponding protein information including amino acid sequences, official gene symbols, UniProt accessions/IDs, chromosome number/ positions, and additional description. The ultimate goal of the ModProt database is to achieve PTM-based diagnosis and personalized medicine through detection of abnormal PTMs by comparing PTM site maps in healthy and disease states using the database.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"88 1","pages":"1-4"},"PeriodicalIF":0.0,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81388188","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A procedure for the direct coupling of affinity chromatography and capillary isoelectric focusing in a single capillary is described. A unified capillary device was made to implement this procedure. The inner surface of a fused-silica capillary was tandemly coated with a chelating polymer, polyiminodiacetate, at the inlet side and with a neutral polymer, polydimethylacrylamide, at the outlet side. After loading a fluorescence-labeled recombinant Fab with a hexahistidine tag, a model sample, the device was rinsed with a high-salt buffer and then with a carrier ampholyte solution to fill the device. The bound Fab was eluted by filling the nickel-chelate column with an anode solution, 100 mM phosphoric acid. A positive voltage was applied at the chelate column side with a pressure at the same side, so as to slightly overwhelm the anodic electroosmosis produced in the acidified chelate column and gradually move the focused protein bands through a stationary fluorescence detector. The mixture of the labeled recombinant Fab at variable concentration and labeled bovine serum albumin at 50 nM was analyzed with the capillary device. A linear relationship between the peak area and the concentration was demonstrated for the Fab at 3.2 pM to 10 nM. The coefficients of variation for the peak area and detection time at 10 nM were 4.3% and 4.4%, respectively. The coupled procedure described here allows total transfer of the specifically adsorbed proteins from the affinity column to the capillary for isoelectric focusing without any compromise in separation efficiency. Removal of excess salts and concentration of dilute samples are also attractive features of the coupled procedure that can provide a new option for the analysis of charge variants of a protein in biological samples.
{"title":"Direct coupling of immobilized metal ion affinity chromatography and capillary isoelectric focusing in a single capillary","authors":"K. Shimura, T. Nagai","doi":"10.2198/JELECTROPH.59.7","DOIUrl":"https://doi.org/10.2198/JELECTROPH.59.7","url":null,"abstract":"A procedure for the direct coupling of affinity chromatography and capillary isoelectric focusing in a single capillary is described. A unified capillary device was made to implement this procedure. The inner surface of a fused-silica capillary was tandemly coated with a chelating polymer, polyiminodiacetate, at the inlet side and with a neutral polymer, polydimethylacrylamide, at the outlet side. After loading a fluorescence-labeled recombinant Fab with a hexahistidine tag, a model sample, the device was rinsed with a high-salt buffer and then with a carrier ampholyte solution to fill the device. The bound Fab was eluted by filling the nickel-chelate column with an anode solution, 100 mM phosphoric acid. A positive voltage was applied at the chelate column side with a pressure at the same side, so as to slightly overwhelm the anodic electroosmosis produced in the acidified chelate column and gradually move the focused protein bands through a stationary fluorescence detector. The mixture of the labeled recombinant Fab at variable concentration and labeled bovine serum albumin at 50 nM was analyzed with the capillary device. A linear relationship between the peak area and the concentration was demonstrated for the Fab at 3.2 pM to 10 nM. The coefficients of variation for the peak area and detection time at 10 nM were 4.3% and 4.4%, respectively. The coupled procedure described here allows total transfer of the specifically adsorbed proteins from the affinity column to the capillary for isoelectric focusing without any compromise in separation efficiency. Removal of excess salts and concentration of dilute samples are also attractive features of the coupled procedure that can provide a new option for the analysis of charge variants of a protein in biological samples.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"137 1","pages":"7-17"},"PeriodicalIF":0.0,"publicationDate":"2015-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76742479","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Takashi Tajima, F. Kito, T. Ohta, K. Shiozawa, A. Kawai, T. Kondo
The potential biological and clinical significance of selenium binding protein 1 (SBP1) has been suggested in various types of cancer. To evaluate the role of SBP1 and reveal the molecular basis for its function, we examined the SBP1 protein complex. A gene transfection assay revealed that overexpression of SBP1 promoted proliferation and migration of A549 lung adenocarcinoma cells. Halo-tag-based affinity purification coupled with liquid chromatography-tandem mass spectrometry identified 23 components of the SBP1 protein complex. The functional classification of these 23 proteins suggests that the SBP1 complex participates in critical biological events including cell structure, protein translation, stress response, chaperone, and apoptosis. Moreover, the SBP1 complex includes several proteins that are aberrantly expressed in cancers. These finding indicate that SBP1 may function coordinately with these multiple proteins to facilitate cancer progression. A comprehensive study of the multiple proteins associated with SPB1 together with an examination of individual proteins will be required to elucidate the roles of aberrant SBP1 regulation in cancer progression.
{"title":"Interactome analysis reveals molecular mechanisms underlying the association between selenium binding protein 1 expression and the malignant features of tumor cells","authors":"Takashi Tajima, F. Kito, T. Ohta, K. Shiozawa, A. Kawai, T. Kondo","doi":"10.2198/JELECTROPH.59.1","DOIUrl":"https://doi.org/10.2198/JELECTROPH.59.1","url":null,"abstract":"The potential biological and clinical significance of selenium binding protein 1 (SBP1) has been suggested in various types of cancer. To evaluate the role of SBP1 and reveal the molecular basis for its function, we examined the SBP1 protein complex. A gene transfection assay revealed that overexpression of SBP1 promoted proliferation and migration of A549 lung adenocarcinoma cells. Halo-tag-based affinity purification coupled with liquid chromatography-tandem mass spectrometry identified 23 components of the SBP1 protein complex. The functional classification of these 23 proteins suggests that the SBP1 complex participates in critical biological events including cell structure, protein translation, stress response, chaperone, and apoptosis. Moreover, the SBP1 complex includes several proteins that are aberrantly expressed in cancers. These finding indicate that SBP1 may function coordinately with these multiple proteins to facilitate cancer progression. A comprehensive study of the multiple proteins associated with SPB1 together with an examination of individual proteins will be required to elucidate the roles of aberrant SBP1 regulation in cancer progression.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"11 1","pages":"1-6"},"PeriodicalIF":0.0,"publicationDate":"2015-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82086173","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
SUMMARY We recently reported a neutral-pH gel system buffered with 2-[bis(2-hydroxyethyl)amino]-2-(hydroxyme-thyl)propane-1,3-diol hydrochloride (Bis-Tris–HCl) for use in Zn 2+ –Phos-tag sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) for advanced profiling of protein phosphorylation. In the current study, we extended the utility of Zn 2+ –Phos-tag SDS-PAGE with the Bis-Tris–HCl buffer system to a detailed analysis of phosphorylated β -catenin, which is closely involved in the ubiquitin-proteasome pathway. The Phos-tag-based approach, followed by Western blotting with an anti- β -catenin antibody, allowed us to assign nine phosphorylated species of β -catenin produced in complicated signaling pathways of cultured HEK293 and SW480 cells. Two-dimensional image coupling with normal Laemmli’s SDS-PAGE as the first dimension gave more detailed information, not only on the phosphorylation of β -catenin, but also on the phosphorylation-dependent polyubiquitination by visualizing multiple ubiquitinated forms derived from two phosphorylated species of β -catenin in lactacystin-treated HEK293 cells. We identified two distinct phosphorylated species of β -catenin that are responsible for polyubiquitination. The first contains phosphorylated residues at S33, S37, T41, and S45, and the second contains these sites and an additional phosphorylated residue at S675. The profiling of double post-translational modifications of β -catenin is consistent with the widely accepted phosphorylation-dependent ubiquitination model in the absence of a Wnt signal.
{"title":"Identification of two phosphorylated species of β-catenin involved in the ubiquitin-proteasome pathway by using two-dimensional Phos-tag affinity electrophoresis","authors":"Emiko Kinoshita-Kikuta, E. Kinoshita, T. Koike","doi":"10.2198/JELECTROPH.58.1","DOIUrl":"https://doi.org/10.2198/JELECTROPH.58.1","url":null,"abstract":"SUMMARY We recently reported a neutral-pH gel system buffered with 2-[bis(2-hydroxyethyl)amino]-2-(hydroxyme-thyl)propane-1,3-diol hydrochloride (Bis-Tris–HCl) for use in Zn 2+ –Phos-tag sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) for advanced profiling of protein phosphorylation. In the current study, we extended the utility of Zn 2+ –Phos-tag SDS-PAGE with the Bis-Tris–HCl buffer system to a detailed analysis of phosphorylated β -catenin, which is closely involved in the ubiquitin-proteasome pathway. The Phos-tag-based approach, followed by Western blotting with an anti- β -catenin antibody, allowed us to assign nine phosphorylated species of β -catenin produced in complicated signaling pathways of cultured HEK293 and SW480 cells. Two-dimensional image coupling with normal Laemmli’s SDS-PAGE as the first dimension gave more detailed information, not only on the phosphorylation of β -catenin, but also on the phosphorylation-dependent polyubiquitination by visualizing multiple ubiquitinated forms derived from two phosphorylated species of β -catenin in lactacystin-treated HEK293 cells. We identified two distinct phosphorylated species of β -catenin that are responsible for polyubiquitination. The first contains phosphorylated residues at S33, S37, T41, and S45, and the second contains these sites and an additional phosphorylated residue at S675. The profiling of double post-translational modifications of β -catenin is consistent with the widely accepted phosphorylation-dependent ubiquitination model in the absence of a Wnt signal.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"96 1","pages":"1-4"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83358515","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Tatsuya Saito, Y. Kawashima, Satoru Minamida, Kazumasa Matsumoto, Keita Araki, T. Matsui, M. Satoh, F. Nomura, M. Iwamura, T. Maeda, S. Baba, Y. Kodera
1 Laboratory of Biomolecular Dynamics, Department of Physics, Kitasato University School of Science 2 Center for Disease Proteomics, Kitasato University School of Science 3 Department of Urology, Kitasato University School of Medicine 4 Division of Natural Products Chemistry, Department of Medicinal Resources, Institute of Natural Medicine, University of Toyama 5 Clinical Proteomics Research Center, Chiba University Hospital 6 Department of Molecular Diagnosis (F8) Graduate School of Medicine, Chiba University
{"title":"Establishment and application of a high-quality comparative analysis strategy for the discovery and small-scale validation of low-abundance biomarker peptides in serum based on an optimized novel peptide extraction method","authors":"Tatsuya Saito, Y. Kawashima, Satoru Minamida, Kazumasa Matsumoto, Keita Araki, T. Matsui, M. Satoh, F. Nomura, M. Iwamura, T. Maeda, S. Baba, Y. Kodera","doi":"10.2198/JELECTROPH.57.1","DOIUrl":"https://doi.org/10.2198/JELECTROPH.57.1","url":null,"abstract":"1 Laboratory of Biomolecular Dynamics, Department of Physics, Kitasato University School of Science 2 Center for Disease Proteomics, Kitasato University School of Science 3 Department of Urology, Kitasato University School of Medicine 4 Division of Natural Products Chemistry, Department of Medicinal Resources, Institute of Natural Medicine, University of Toyama 5 Clinical Proteomics Research Center, Chiba University Hospital 6 Department of Molecular Diagnosis (F8) Graduate School of Medicine, Chiba University","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"33 1","pages":"1-9"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74795568","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2012-01-01DOI: 10.2198/JELECTROPH.56.19
M. Negoro, Mina Sawano, Hiromi Sawamura, S. Ebara, Toshiaki Watanabe
{"title":"Analysis of glucose-protein interaction using p-hydroxyacetophenone-Sepharose affinity resin —Glucose decreases alcohol dehydrogenase activity in vitro—","authors":"M. Negoro, Mina Sawano, Hiromi Sawamura, S. Ebara, Toshiaki Watanabe","doi":"10.2198/JELECTROPH.56.19","DOIUrl":"https://doi.org/10.2198/JELECTROPH.56.19","url":null,"abstract":"","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"6 1","pages":"19-24"},"PeriodicalIF":0.0,"publicationDate":"2012-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76087821","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nobue Sakai, R. Kubota, K. Shiba, Y. Hattori, S. Iijima
Urinary albumin (ALB) and transferrin (TF) are useful markers for impaired renal function. Compared to ALB, TF is more readily excreted into urine from an early stage of renal failure because of its smaller number of negative charges. However, few evidence of a change in isoelectric point (pI) value of urinary TF in nephropathy patients has been reported. We analyzed pI values of urinary TF by an isoelectric focusing, and compared the results in normal subjects with those in diabetic patients at various stages of nephropathy. Urine samples of diabetics were randomly collected from 24 patients and 7 healthy volunteers. An apotransferrin with a high pI value was detected in samples of healthy subjects. However, the pI value of TF bands derived from diabetic patients shifted to the lower side, and the more nephropathy stages progressed, greater change of pI value was detected. In addition, TF with low iron-binding capacity was detected in some samples from diabetics. These results suggest that determination of the pI value of urinary TF in diabetic patients may be useful diagnostic markers for an early detection of diabetic nephropathy.
{"title":"Isoelectric focusing of urinary transferrin for detection of early stages of diabetic nephropathy","authors":"Nobue Sakai, R. Kubota, K. Shiba, Y. Hattori, S. Iijima","doi":"10.2198/JELECTROPH.56.1","DOIUrl":"https://doi.org/10.2198/JELECTROPH.56.1","url":null,"abstract":"Urinary albumin (ALB) and transferrin (TF) are useful markers for impaired renal function. Compared to ALB, TF is more readily excreted into urine from an early stage of renal failure because of its smaller number of negative charges. However, few evidence of a change in isoelectric point (pI) value of urinary TF in nephropathy patients has been reported. We analyzed pI values of urinary TF by an isoelectric focusing, and compared the results in normal subjects with those in diabetic patients at various stages of nephropathy. Urine samples of diabetics were randomly collected from 24 patients and 7 healthy volunteers. An apotransferrin with a high pI value was detected in samples of healthy subjects. However, the pI value of TF bands derived from diabetic patients shifted to the lower side, and the more nephropathy stages progressed, greater change of pI value was detected. In addition, TF with low iron-binding capacity was detected in some samples from diabetics. These results suggest that determination of the pI value of urinary TF in diabetic patients may be useful diagnostic markers for an early detection of diabetic nephropathy.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"37 1","pages":"1-6"},"PeriodicalIF":0.0,"publicationDate":"2012-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74045227","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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