Detection of Differentially Expressed Basal Cell Proteins by Mass Spectrometry
The ability of cells to modulate interactions with each other and the substrate is essential for epithelial tissue remodeling during processes such as wound healing and tumor progression. However, despite strides made in the field of proteomics, proteins involved in adhesion have been difficult to s...
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Veröffentlicht in: | Molecular & cellular proteomics 2010-02, Vol.9 (2), p.351-361 |
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creator | Todorović, Viktor Desai, Bhushan V. Eigenheer, Richard A. Yin, Taofei Amargo, Evangeline V. Mrksich, Milan Green, Kathleen J. Patterson, Melanie J. Schroeder |
description | The ability of cells to modulate interactions with each other and the substrate is essential for epithelial tissue remodeling during processes such as wound healing and tumor progression. However, despite strides made in the field of proteomics, proteins involved in adhesion have been difficult to study. Here, we report a method for the enrichment and analysis of proteins associated with the basal surface of the cell and its underlying matrix. The enrichment involves deroofing the cells with 20 mm ammonium hydroxide and the removal of cytosolic and organellar proteins by stringent water wash. Proteomic profiling was achieved by LC-FTMS, which allowed comparison of differentially expressed or shared proteins under different cell states. First, we analyzed and compared the basal cell components of mouse keratinocytes lacking the cell-cell junction molecule plakoglobin with their control counterparts. Changes in the molecules involved in motility and invasion were detected in plakoglobin-deficient cells, including decreased detection of fibronectin, integrin β4, and FAT tumor suppressor. Second, we assessed the differences in basal cell components between two human oral squamous cell carcinoma lines originating from different sites in the oral cavity (CAL33 and UM-SCC-1). The data show differences between the two lines in the type and abundance of proteins specific to cell adhesion, migration, and angiogenesis. Therefore, the method described here has the potential to serve as a platform to assess proteomic changes in basal cell components including extracellular and adhesion-specific proteins involved in wound healing, cancer, and chronic and acquired adhesion-related disorders. |
doi_str_mv | 10.1074/mcp.M900358-MCP200 |
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Schroeder</creator><creatorcontrib>Todorović, Viktor ; Desai, Bhushan V. ; Eigenheer, Richard A. ; Yin, Taofei ; Amargo, Evangeline V. ; Mrksich, Milan ; Green, Kathleen J. ; Patterson, Melanie J. Schroeder</creatorcontrib><description>The ability of cells to modulate interactions with each other and the substrate is essential for epithelial tissue remodeling during processes such as wound healing and tumor progression. However, despite strides made in the field of proteomics, proteins involved in adhesion have been difficult to study. Here, we report a method for the enrichment and analysis of proteins associated with the basal surface of the cell and its underlying matrix. The enrichment involves deroofing the cells with 20 mm ammonium hydroxide and the removal of cytosolic and organellar proteins by stringent water wash. Proteomic profiling was achieved by LC-FTMS, which allowed comparison of differentially expressed or shared proteins under different cell states. First, we analyzed and compared the basal cell components of mouse keratinocytes lacking the cell-cell junction molecule plakoglobin with their control counterparts. Changes in the molecules involved in motility and invasion were detected in plakoglobin-deficient cells, including decreased detection of fibronectin, integrin β4, and FAT tumor suppressor. Second, we assessed the differences in basal cell components between two human oral squamous cell carcinoma lines originating from different sites in the oral cavity (CAL33 and UM-SCC-1). The data show differences between the two lines in the type and abundance of proteins specific to cell adhesion, migration, and angiogenesis. Therefore, the method described here has the potential to serve as a platform to assess proteomic changes in basal cell components including extracellular and adhesion-specific proteins involved in wound healing, cancer, and chronic and acquired adhesion-related disorders.</description><identifier>ISSN: 1535-9476</identifier><identifier>EISSN: 1535-9484</identifier><identifier>DOI: 10.1074/mcp.M900358-MCP200</identifier><identifier>PMID: 19955077</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Ammonium Hydroxide ; Animals ; Carcinoma, Squamous Cell - metabolism ; Carcinoma, Squamous Cell - pathology ; Cell Adhesion - drug effects ; Cell Line, Tumor ; Cell Membrane - drug effects ; Cell Membrane - metabolism ; Extracellular Matrix - drug effects ; Extracellular Matrix - metabolism ; gamma Catenin - deficiency ; gamma Catenin - metabolism ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; Hydroxides - pharmacology ; Keratinocytes - cytology ; Keratinocytes - drug effects ; Keratinocytes - metabolism ; Mass Spectrometry - methods ; Mice ; Mouth Neoplasms - metabolism ; Mouth Neoplasms - pathology ; Peptides - chemistry ; Peptides - metabolism ; Proteins - genetics ; Proteins - metabolism</subject><ispartof>Molecular & cellular proteomics, 2010-02, Vol.9 (2), p.351-361</ispartof><rights>2010 © 2010 ASBMB. 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Schroeder</creatorcontrib><title>Detection of Differentially Expressed Basal Cell Proteins by Mass Spectrometry</title><title>Molecular & cellular proteomics</title><addtitle>Mol Cell Proteomics</addtitle><description>The ability of cells to modulate interactions with each other and the substrate is essential for epithelial tissue remodeling during processes such as wound healing and tumor progression. However, despite strides made in the field of proteomics, proteins involved in adhesion have been difficult to study. Here, we report a method for the enrichment and analysis of proteins associated with the basal surface of the cell and its underlying matrix. The enrichment involves deroofing the cells with 20 mm ammonium hydroxide and the removal of cytosolic and organellar proteins by stringent water wash. Proteomic profiling was achieved by LC-FTMS, which allowed comparison of differentially expressed or shared proteins under different cell states. First, we analyzed and compared the basal cell components of mouse keratinocytes lacking the cell-cell junction molecule plakoglobin with their control counterparts. Changes in the molecules involved in motility and invasion were detected in plakoglobin-deficient cells, including decreased detection of fibronectin, integrin β4, and FAT tumor suppressor. Second, we assessed the differences in basal cell components between two human oral squamous cell carcinoma lines originating from different sites in the oral cavity (CAL33 and UM-SCC-1). The data show differences between the two lines in the type and abundance of proteins specific to cell adhesion, migration, and angiogenesis. Therefore, the method described here has the potential to serve as a platform to assess proteomic changes in basal cell components including extracellular and adhesion-specific proteins involved in wound healing, cancer, and chronic and acquired adhesion-related disorders.</description><subject>Ammonium Hydroxide</subject><subject>Animals</subject><subject>Carcinoma, Squamous Cell - metabolism</subject><subject>Carcinoma, Squamous Cell - pathology</subject><subject>Cell Adhesion - drug effects</subject><subject>Cell Line, Tumor</subject><subject>Cell Membrane - drug effects</subject><subject>Cell Membrane - metabolism</subject><subject>Extracellular Matrix - drug effects</subject><subject>Extracellular Matrix - metabolism</subject><subject>gamma Catenin - deficiency</subject><subject>gamma Catenin - metabolism</subject><subject>Gene Expression Profiling</subject><subject>Gene Expression Regulation</subject><subject>Humans</subject><subject>Hydroxides - pharmacology</subject><subject>Keratinocytes - cytology</subject><subject>Keratinocytes - drug effects</subject><subject>Keratinocytes - metabolism</subject><subject>Mass Spectrometry - methods</subject><subject>Mice</subject><subject>Mouth Neoplasms - metabolism</subject><subject>Mouth Neoplasms - pathology</subject><subject>Peptides - chemistry</subject><subject>Peptides - metabolism</subject><subject>Proteins - genetics</subject><subject>Proteins - metabolism</subject><issn>1535-9476</issn><issn>1535-9484</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kN1u1DAQRi0EoqXwAlyAXyBlnNiJLSEk2JYfqQuVgGvLdsZdoySO7NB2375GWRW46YVlSz7fN6NDyEsGpww6_mZ08-lWATRCVtvNZQ3wiBwz0YhKcckf37-79og8y_kXQA2sE0_JEVNKCOi6Y_L1DBd0S4gTjZ6eBe8x4bQEMwx7en47J8wZe_rBZDPQDQ4DvUxxwTBlavd0a3Km3-dSkOKIS9o_J0-8GTK-ONwn5OfH8x-bz9XFt09fNu8vKsc7uVTIlXJdr6RizEphW9e3XLZ9zzpwgnFgUI73rbBe1EZZC8Bdjc75VoJVzQl5t_bOv-2IvSsrJzPoOYXRpL2OJuj_f6aw01fxWteyAclFKajXApdizgn9fZaB_mNXF7v6YFevdkvo1b9T_0YOOgvwegV24Wp3ExJqG6Lb4aiVrnUjWCHergQWO9cBk84u4OSwL7RbdB_DQxvcAe2al2A</recordid><startdate>20100201</startdate><enddate>20100201</enddate><creator>Todorović, Viktor</creator><creator>Desai, Bhushan V.</creator><creator>Eigenheer, Richard A.</creator><creator>Yin, Taofei</creator><creator>Amargo, Evangeline V.</creator><creator>Mrksich, Milan</creator><creator>Green, Kathleen J.</creator><creator>Patterson, Melanie J. Schroeder</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><general>The American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</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>5PM</scope></search><sort><creationdate>20100201</creationdate><title>Detection of Differentially Expressed Basal Cell Proteins by Mass Spectrometry</title><author>Todorović, Viktor ; Desai, Bhushan V. ; Eigenheer, Richard A. ; Yin, Taofei ; Amargo, Evangeline V. ; Mrksich, Milan ; Green, Kathleen J. ; Patterson, Melanie J. 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The enrichment involves deroofing the cells with 20 mm ammonium hydroxide and the removal of cytosolic and organellar proteins by stringent water wash. Proteomic profiling was achieved by LC-FTMS, which allowed comparison of differentially expressed or shared proteins under different cell states. First, we analyzed and compared the basal cell components of mouse keratinocytes lacking the cell-cell junction molecule plakoglobin with their control counterparts. Changes in the molecules involved in motility and invasion were detected in plakoglobin-deficient cells, including decreased detection of fibronectin, integrin β4, and FAT tumor suppressor. Second, we assessed the differences in basal cell components between two human oral squamous cell carcinoma lines originating from different sites in the oral cavity (CAL33 and UM-SCC-1). The data show differences between the two lines in the type and abundance of proteins specific to cell adhesion, migration, and angiogenesis. 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subjects | Ammonium Hydroxide Animals Carcinoma, Squamous Cell - metabolism Carcinoma, Squamous Cell - pathology Cell Adhesion - drug effects Cell Line, Tumor Cell Membrane - drug effects Cell Membrane - metabolism Extracellular Matrix - drug effects Extracellular Matrix - metabolism gamma Catenin - deficiency gamma Catenin - metabolism Gene Expression Profiling Gene Expression Regulation Humans Hydroxides - pharmacology Keratinocytes - cytology Keratinocytes - drug effects Keratinocytes - metabolism Mass Spectrometry - methods Mice Mouth Neoplasms - metabolism Mouth Neoplasms - pathology Peptides - chemistry Peptides - metabolism Proteins - genetics Proteins - metabolism |
title | Detection of Differentially Expressed Basal Cell Proteins by Mass Spectrometry |
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