Proteome analysis of differentiating human myoblasts by dialysis-assisted two-dimensional gel electrophoresis (DAGE)
In the present study, modifications in cytosolic expressed proteins during human myoblast differentiation were studied by dialysis-assisted 2-DE (DAGE, [1]). About 1000 spots were analysed on the 5th and 13th day of differentiation with a dynamic range of protein expression exceeding 1000-fold. Duri...
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| Veröffentlicht in: | Proteomics (Weinheim) 2008-01, Vol.8 (2), p.264-278 |
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| creator | Gonnet, Florence Bouazza, Belaid Millot, Gaël Armel Ziaei, Simin Garcia, Luis Butler-Browne, Gillian S Mouly, Vincent Tortajada, Jeanine Danos, Olivier Svinartchouk, Fédor |
| description | In the present study, modifications in cytosolic expressed proteins during human myoblast differentiation were studied by dialysis-assisted 2-DE (DAGE, [1]). About 1000 spots were analysed on the 5th and 13th day of differentiation with a dynamic range of protein expression exceeding 1000-fold. During myogenic differentiation, the number of nonmatching spots as well as the extent of quantitative differences between matched spots significantly increased. Over one hundred differentially expressed spots were excised and identified by MALDI-TOF MS. The differentiation-associated expression pattern of eight proteins was validated by Western blot analysis. Differential expression of several proteins was demonstrated for the first time in human myotubes. Interestingly, Ingenuity pathway analysis grouped 30 of these proteins into two overlapping networks containing as principal nodes IGF-1 and tumour necrosis factor, two proteins known to play a crucial role in cytogenesis. Our results illustrate the large rearrangement of the proteome during the differentiation of human myoblasts and provide evidence for new partners involved in this complex process. |
| doi_str_mv | 10.1002/pmic.200700261 |
| format | Article |
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About 1000 spots were analysed on the 5th and 13th day of differentiation with a dynamic range of protein expression exceeding 1000-fold. During myogenic differentiation, the number of nonmatching spots as well as the extent of quantitative differences between matched spots significantly increased. Over one hundred differentially expressed spots were excised and identified by MALDI-TOF MS. The differentiation-associated expression pattern of eight proteins was validated by Western blot analysis. Differential expression of several proteins was demonstrated for the first time in human myotubes. Interestingly, Ingenuity pathway analysis grouped 30 of these proteins into two overlapping networks containing as principal nodes IGF-1 and tumour necrosis factor, two proteins known to play a crucial role in cytogenesis. Our results illustrate the large rearrangement of the proteome during the differentiation of human myoblasts and provide evidence for new partners involved in this complex process.</description><identifier>ISSN: 1615-9853</identifier><identifier>EISSN: 1615-9861</identifier><identifier>DOI: 10.1002/pmic.200700261</identifier><identifier>PMID: 18203276</identifier><language>eng</language><publisher>Weinheim: Wiley-VCH Verlag</publisher><subject>2-D PAGE ; Analytical, structural and metabolic biochemistry ; Biochemistry, Molecular Biology ; Biological and medical sciences ; Blotting, Western ; Cell Differentiation ; Cytosol - chemistry ; Dialysis - methods ; Differential expression ; Electrophoresis, Gel, Two-Dimensional - methods ; Factor XIII - analysis ; Fundamental and applied biological sciences. Psychology ; Genomics ; Guanine Nucleotide Dissociation Inhibitors - analysis ; Heterogeneous-Nuclear Ribonucleoprotein K - analysis ; Human ; Humans ; Life Sciences ; Mass spectrometry ; Miscellaneous ; Myoblasts - chemistry ; Protein identification ; Proteins ; Proteomics - methods ; rho-Specific Guanine Nucleotide Dissociation Inhibitors ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; STAT1 Transcription Factor - analysis ; Stathmin - analysis</subject><ispartof>Proteomics (Weinheim), 2008-01, Vol.8 (2), p.264-278</ispartof><rights>Copyright © 2008 WILEY‐VCH Verlag GmbH & Co. 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About 1000 spots were analysed on the 5th and 13th day of differentiation with a dynamic range of protein expression exceeding 1000-fold. During myogenic differentiation, the number of nonmatching spots as well as the extent of quantitative differences between matched spots significantly increased. Over one hundred differentially expressed spots were excised and identified by MALDI-TOF MS. The differentiation-associated expression pattern of eight proteins was validated by Western blot analysis. Differential expression of several proteins was demonstrated for the first time in human myotubes. Interestingly, Ingenuity pathway analysis grouped 30 of these proteins into two overlapping networks containing as principal nodes IGF-1 and tumour necrosis factor, two proteins known to play a crucial role in cytogenesis. Our results illustrate the large rearrangement of the proteome during the differentiation of human myoblasts and provide evidence for new partners involved in this complex process.</description><subject>2-D PAGE</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Biochemistry, Molecular Biology</subject><subject>Biological and medical sciences</subject><subject>Blotting, Western</subject><subject>Cell Differentiation</subject><subject>Cytosol - chemistry</subject><subject>Dialysis - methods</subject><subject>Differential expression</subject><subject>Electrophoresis, Gel, Two-Dimensional - methods</subject><subject>Factor XIII - analysis</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genomics</subject><subject>Guanine Nucleotide Dissociation Inhibitors - analysis</subject><subject>Heterogeneous-Nuclear Ribonucleoprotein K - analysis</subject><subject>Human</subject><subject>Humans</subject><subject>Life Sciences</subject><subject>Mass spectrometry</subject><subject>Miscellaneous</subject><subject>Myoblasts - chemistry</subject><subject>Protein identification</subject><subject>Proteins</subject><subject>Proteomics - methods</subject><subject>rho-Specific Guanine Nucleotide Dissociation Inhibitors</subject><subject>Sensitivity and Specificity</subject><subject>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</subject><subject>STAT1 Transcription Factor - analysis</subject><subject>Stathmin - analysis</subject><issn>1615-9853</issn><issn>1615-9861</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc9v0zAUxyMEYqNw5Qi5gNghxc-O7fRYla2bKDANph4tJ3lpDUlc7JSR_x5HqQI3Lv6lz_v46X2j6CWQORBC3x8aU8wpITJcBDyKzkEATxaZgMfTmbOz6Jn33wkBmS3k0-gMMkoYleI86m6d7dA2GOtW1703PrZVXJqqQodtZ3Rn2l28Pza6jZve5rX2nY_zPiAjnmgf1g7LuHuwSWkabL2xwRXvsI6xxqJz9rC3Dgf3uw_L9eXF8-hJpWuPL077LLq_uvy2uk42X9Y3q-UmKTjjkAAglgJYKnSelgTDUSDQqpKEpzIvRZ6hRk4BgGKxoIxLSUWVl8jSstDAZtHF6N3rWh2cabTrldVGXS83angLQ5PAIPs1sG9H9uDszyP6TjXGF1jXukV79EqS4Icwy1k0H8HCWe8dVpMZiBoyUUMmasokFLw6mY95g-Vf_BRCAN6cAO0LXVdOt4XxExdMPAUxcIuRezA19v_5Vt1-uln920Qy1g5Z_Z5qtfuhhGSSq-3ntfp4t91cbdM7tQ3865GvtFV650I_918pAUZIxlPKCfsDgIy-3g</recordid><startdate>20080101</startdate><enddate>20080101</enddate><creator>Gonnet, Florence</creator><creator>Bouazza, Belaid</creator><creator>Millot, Gaël Armel</creator><creator>Ziaei, Simin</creator><creator>Garcia, Luis</creator><creator>Butler-Browne, Gillian S</creator><creator>Mouly, Vincent</creator><creator>Tortajada, Jeanine</creator><creator>Danos, Olivier</creator><creator>Svinartchouk, Fédor</creator><general>Wiley-VCH Verlag</general><general>WILEY-VCH Verlag</general><general>WILEY‐VCH Verlag</general><general>Wiley-VCH</general><scope>FBQ</scope><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>1XC</scope><orcidid>https://orcid.org/0000-0002-0591-3509</orcidid><orcidid>https://orcid.org/0000-0001-8003-198X</orcidid><orcidid>https://orcid.org/0000-0003-3809-4033</orcidid></search><sort><creationdate>20080101</creationdate><title>Proteome analysis of differentiating human myoblasts by dialysis-assisted two-dimensional gel electrophoresis (DAGE)</title><author>Gonnet, Florence ; Bouazza, Belaid ; Millot, Gaël Armel ; Ziaei, Simin ; Garcia, Luis ; Butler-Browne, Gillian S ; Mouly, Vincent ; Tortajada, Jeanine ; Danos, Olivier ; Svinartchouk, Fédor</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5351-11eed61346ab4d0e6136e12ff70547bd6b8eae521112ec92357726fbde34dca13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>2-D PAGE</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Biochemistry, Molecular Biology</topic><topic>Biological and medical sciences</topic><topic>Blotting, Western</topic><topic>Cell Differentiation</topic><topic>Cytosol - chemistry</topic><topic>Dialysis - methods</topic><topic>Differential expression</topic><topic>Electrophoresis, Gel, Two-Dimensional - methods</topic><topic>Factor XIII - analysis</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genomics</topic><topic>Guanine Nucleotide Dissociation Inhibitors - analysis</topic><topic>Heterogeneous-Nuclear Ribonucleoprotein K - analysis</topic><topic>Human</topic><topic>Humans</topic><topic>Life Sciences</topic><topic>Mass spectrometry</topic><topic>Miscellaneous</topic><topic>Myoblasts - chemistry</topic><topic>Protein identification</topic><topic>Proteins</topic><topic>Proteomics - methods</topic><topic>rho-Specific Guanine Nucleotide Dissociation Inhibitors</topic><topic>Sensitivity and Specificity</topic><topic>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</topic><topic>STAT1 Transcription Factor - analysis</topic><topic>Stathmin - analysis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gonnet, Florence</creatorcontrib><creatorcontrib>Bouazza, Belaid</creatorcontrib><creatorcontrib>Millot, Gaël Armel</creatorcontrib><creatorcontrib>Ziaei, Simin</creatorcontrib><creatorcontrib>Garcia, Luis</creatorcontrib><creatorcontrib>Butler-Browne, Gillian S</creatorcontrib><creatorcontrib>Mouly, Vincent</creatorcontrib><creatorcontrib>Tortajada, Jeanine</creatorcontrib><creatorcontrib>Danos, Olivier</creatorcontrib><creatorcontrib>Svinartchouk, Fédor</creatorcontrib><collection>AGRIS</collection><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Hyper Article en Ligne (HAL)</collection><jtitle>Proteomics (Weinheim)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gonnet, Florence</au><au>Bouazza, Belaid</au><au>Millot, Gaël Armel</au><au>Ziaei, Simin</au><au>Garcia, Luis</au><au>Butler-Browne, Gillian S</au><au>Mouly, Vincent</au><au>Tortajada, Jeanine</au><au>Danos, Olivier</au><au>Svinartchouk, Fédor</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Proteome analysis of differentiating human myoblasts by dialysis-assisted two-dimensional gel electrophoresis (DAGE)</atitle><jtitle>Proteomics (Weinheim)</jtitle><addtitle>Proteomics</addtitle><date>2008-01-01</date><risdate>2008</risdate><volume>8</volume><issue>2</issue><spage>264</spage><epage>278</epage><pages>264-278</pages><issn>1615-9853</issn><eissn>1615-9861</eissn><abstract>In the present study, modifications in cytosolic expressed proteins during human myoblast differentiation were studied by dialysis-assisted 2-DE (DAGE, [1]). About 1000 spots were analysed on the 5th and 13th day of differentiation with a dynamic range of protein expression exceeding 1000-fold. During myogenic differentiation, the number of nonmatching spots as well as the extent of quantitative differences between matched spots significantly increased. Over one hundred differentially expressed spots were excised and identified by MALDI-TOF MS. The differentiation-associated expression pattern of eight proteins was validated by Western blot analysis. Differential expression of several proteins was demonstrated for the first time in human myotubes. Interestingly, Ingenuity pathway analysis grouped 30 of these proteins into two overlapping networks containing as principal nodes IGF-1 and tumour necrosis factor, two proteins known to play a crucial role in cytogenesis. Our results illustrate the large rearrangement of the proteome during the differentiation of human myoblasts and provide evidence for new partners involved in this complex process.</abstract><cop>Weinheim</cop><pub>Wiley-VCH Verlag</pub><pmid>18203276</pmid><doi>10.1002/pmic.200700261</doi><tpages>15</tpages><orcidid>https://orcid.org/0000-0002-0591-3509</orcidid><orcidid>https://orcid.org/0000-0001-8003-198X</orcidid><orcidid>https://orcid.org/0000-0003-3809-4033</orcidid></addata></record> |
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| subjects | 2-D PAGE Analytical, structural and metabolic biochemistry Biochemistry, Molecular Biology Biological and medical sciences Blotting, Western Cell Differentiation Cytosol - chemistry Dialysis - methods Differential expression Electrophoresis, Gel, Two-Dimensional - methods Factor XIII - analysis Fundamental and applied biological sciences. Psychology Genomics Guanine Nucleotide Dissociation Inhibitors - analysis Heterogeneous-Nuclear Ribonucleoprotein K - analysis Human Humans Life Sciences Mass spectrometry Miscellaneous Myoblasts - chemistry Protein identification Proteins Proteomics - methods rho-Specific Guanine Nucleotide Dissociation Inhibitors Sensitivity and Specificity Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization STAT1 Transcription Factor - analysis Stathmin - analysis |
| title | Proteome analysis of differentiating human myoblasts by dialysis-assisted two-dimensional gel electrophoresis (DAGE) |
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