Direct on-membrane peptide mass fingerprinting with MALDI–MS of tyrosine-phosphorylated proteins detected by immunostaining
We have identified tyrosine-phosphorylated proteins on membrane from A-431 human epidermoid carcinoma cells by using detection with anti-phosphotyrosine antibody followed by PMF analysis. In there, on-membrane digestion for these protein spots was carried out on microscale region using chemical inkj...
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| Veröffentlicht in: | Journal of chromatography. B 2007-02, Vol.847 (1), p.24-29 |
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| container_title | Journal of chromatography. B |
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| creator | Nakanishi, Tsuyoshi Ando, Eiji Furuta, Masaru Tsunasawa, Susumu Nishimura, Osamu |
| description | We have identified tyrosine-phosphorylated proteins on membrane from A-431 human epidermoid carcinoma cells by using detection with anti-phosphotyrosine antibody followed by PMF analysis. In there, on-membrane digestion for these protein spots was carried out on microscale region using chemical inkjet technology and the resulting tryptic digests were directly analyzed by MALDI–TOF MS. Proteins identified by a database search included phosphoproteins that are known to be markedly phosphorylated on tyrosine sites after the cells are treated with epidermal growth factor (EGF). This procedure is a rapid and easily handled approach that enables both detection and identification of phosphoproteins on a single blot membrane. |
| doi_str_mv | 10.1016/j.jchromb.2006.08.024 |
| format | Article |
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In there, on-membrane digestion for these protein spots was carried out on microscale region using chemical inkjet technology and the resulting tryptic digests were directly analyzed by MALDI–TOF MS. Proteins identified by a database search included phosphoproteins that are known to be markedly phosphorylated on tyrosine sites after the cells are treated with epidermal growth factor (EGF). This procedure is a rapid and easily handled approach that enables both detection and identification of phosphoproteins on a single blot membrane.</description><identifier>ISSN: 1570-0232</identifier><identifier>EISSN: 1873-376X</identifier><identifier>DOI: 10.1016/j.jchromb.2006.08.024</identifier><identifier>PMID: 16959554</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Analysis ; Analytical, structural and metabolic biochemistry ; Antibodies, Phospho-Specific - analysis ; Biological and medical sciences ; Blotting, Western - methods ; Carcinoma, Squamous Cell ; Cell Extracts - chemistry ; Cell Line, Tumor - chemistry ; Cell Line, Tumor - drug effects ; Cell Membrane - chemistry ; Electrophoresis, Gel, Two-Dimensional - methods ; Epidermal Growth Factor - pharmacology ; Fundamental and applied biological sciences. Psychology ; General pharmacology ; Humans ; Immunodetection ; MALDI–TOF MS ; Medical sciences ; Molecular Weight ; On-membrane digestion ; Peptide Mapping ; Pharmacology. Drug treatments ; Phosphoproteins - analysis ; Phosphoproteins - immunology ; Phosphorylation ; Phosphotyrosine - analysis ; Phosphotyrosine - immunology ; Protein identification ; Proteins - isolation & purification ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods ; Tandem Mass Spectrometry - methods</subject><ispartof>Journal of chromatography. 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B</title><addtitle>J Chromatogr B Analyt Technol Biomed Life Sci</addtitle><description>We have identified tyrosine-phosphorylated proteins on membrane from A-431 human epidermoid carcinoma cells by using detection with anti-phosphotyrosine antibody followed by PMF analysis. In there, on-membrane digestion for these protein spots was carried out on microscale region using chemical inkjet technology and the resulting tryptic digests were directly analyzed by MALDI–TOF MS. Proteins identified by a database search included phosphoproteins that are known to be markedly phosphorylated on tyrosine sites after the cells are treated with epidermal growth factor (EGF). This procedure is a rapid and easily handled approach that enables both detection and identification of phosphoproteins on a single blot membrane.</description><subject>Analysis</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Antibodies, Phospho-Specific - analysis</subject><subject>Biological and medical sciences</subject><subject>Blotting, Western - methods</subject><subject>Carcinoma, Squamous Cell</subject><subject>Cell Extracts - chemistry</subject><subject>Cell Line, Tumor - chemistry</subject><subject>Cell Line, Tumor - drug effects</subject><subject>Cell Membrane - chemistry</subject><subject>Electrophoresis, Gel, Two-Dimensional - methods</subject><subject>Epidermal Growth Factor - pharmacology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>General pharmacology</subject><subject>Humans</subject><subject>Immunodetection</subject><subject>MALDI–TOF MS</subject><subject>Medical sciences</subject><subject>Molecular Weight</subject><subject>On-membrane digestion</subject><subject>Peptide Mapping</subject><subject>Pharmacology. Drug treatments</subject><subject>Phosphoproteins - analysis</subject><subject>Phosphoproteins - immunology</subject><subject>Phosphorylation</subject><subject>Phosphotyrosine - analysis</subject><subject>Phosphotyrosine - immunology</subject><subject>Protein identification</subject><subject>Proteins - isolation & purification</subject><subject>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods</subject><subject>Tandem Mass Spectrometry - methods</subject><issn>1570-0232</issn><issn>1873-376X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkctu1DAUQC0EoqXwCSBvYJdwHcdxskJVW6DSVCwAiZ3lxw3jUV7YnqJZIPUf-EO-BJeJ1CULy1fWua9jQl4yKBmw5u2u3NltmEdTVgBNCW0JVf2InLJW8oLL5tvjHAsJBVS8OiHPYtwBMAmSPyUnrOlEJ0R9Sn5d-oA20XkqRhxN0BPSBZfkHdJRx0h7P33HsAQ_pRzRnz5t6c355vL6z93vm8907mk6hDn6CYtlO8d8wmHQCR1dwpzQT5E6TLlFfjEH6sdxP80xaT_lcs_Jk14PEV-s9xn5-v7qy8XHYvPpw_XF-aawdVWlQpqqMsjy7A4Fx85wYA6tZdxVWkNnetTOGYlMNi2XPTgjbNfUthatsV3Pz8ibY9080489xqRGHy0OQ1533kfVdADAG5ZBcQRt3ikG7FXefNThoBioe-9qp1bv6t67glZl7znv1dpgb0Z0D1mr6Ay8XgEdrR76LNr6-MC1opZdKzL37shh1nHrMahoPU4W3b9_Um72_xnlL1LTqJo</recordid><startdate>20070215</startdate><enddate>20070215</enddate><creator>Nakanishi, Tsuyoshi</creator><creator>Ando, Eiji</creator><creator>Furuta, Masaru</creator><creator>Tsunasawa, Susumu</creator><creator>Nishimura, Osamu</creator><general>Elsevier B.V</general><general>Elsevier Science</general><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></search><sort><creationdate>20070215</creationdate><title>Direct on-membrane peptide mass fingerprinting with MALDI–MS of tyrosine-phosphorylated proteins detected by immunostaining</title><author>Nakanishi, Tsuyoshi ; Ando, Eiji ; Furuta, Masaru ; Tsunasawa, Susumu ; Nishimura, Osamu</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c422t-7b22be1073de53e9b301decc13d2aa09bfeaddb7e176837f0db5c964c458bc9f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Analysis</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Antibodies, Phospho-Specific - analysis</topic><topic>Biological and medical sciences</topic><topic>Blotting, Western - methods</topic><topic>Carcinoma, Squamous Cell</topic><topic>Cell Extracts - chemistry</topic><topic>Cell Line, Tumor - chemistry</topic><topic>Cell Line, Tumor - drug effects</topic><topic>Cell Membrane - chemistry</topic><topic>Electrophoresis, Gel, Two-Dimensional - methods</topic><topic>Epidermal Growth Factor - pharmacology</topic><topic>Fundamental and applied biological sciences. 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Drug treatments</topic><topic>Phosphoproteins - analysis</topic><topic>Phosphoproteins - immunology</topic><topic>Phosphorylation</topic><topic>Phosphotyrosine - analysis</topic><topic>Phosphotyrosine - immunology</topic><topic>Protein identification</topic><topic>Proteins - isolation & purification</topic><topic>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods</topic><topic>Tandem Mass Spectrometry - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nakanishi, Tsuyoshi</creatorcontrib><creatorcontrib>Ando, Eiji</creatorcontrib><creatorcontrib>Furuta, Masaru</creatorcontrib><creatorcontrib>Tsunasawa, Susumu</creatorcontrib><creatorcontrib>Nishimura, Osamu</creatorcontrib><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><jtitle>Journal of chromatography. 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In there, on-membrane digestion for these protein spots was carried out on microscale region using chemical inkjet technology and the resulting tryptic digests were directly analyzed by MALDI–TOF MS. Proteins identified by a database search included phosphoproteins that are known to be markedly phosphorylated on tyrosine sites after the cells are treated with epidermal growth factor (EGF). This procedure is a rapid and easily handled approach that enables both detection and identification of phosphoproteins on a single blot membrane.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>16959554</pmid><doi>10.1016/j.jchromb.2006.08.024</doi><tpages>6</tpages></addata></record> |
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| subjects | Analysis Analytical, structural and metabolic biochemistry Antibodies, Phospho-Specific - analysis Biological and medical sciences Blotting, Western - methods Carcinoma, Squamous Cell Cell Extracts - chemistry Cell Line, Tumor - chemistry Cell Line, Tumor - drug effects Cell Membrane - chemistry Electrophoresis, Gel, Two-Dimensional - methods Epidermal Growth Factor - pharmacology Fundamental and applied biological sciences. Psychology General pharmacology Humans Immunodetection MALDI–TOF MS Medical sciences Molecular Weight On-membrane digestion Peptide Mapping Pharmacology. Drug treatments Phosphoproteins - analysis Phosphoproteins - immunology Phosphorylation Phosphotyrosine - analysis Phosphotyrosine - immunology Protein identification Proteins - isolation & purification Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods Tandem Mass Spectrometry - methods |
| title | Direct on-membrane peptide mass fingerprinting with MALDI–MS of tyrosine-phosphorylated proteins detected by immunostaining |
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