High-throughput analysis of GST-fusion protein expression and activity-dependent protein interactions on GST-fusion protein arrays with a spectral surface plasmon resonance biosensor
We modified gold arrays with a glutathione (GSH) surface, and investigated high‐throughput protein interactions with a spectral surface plasmon resonance (SPR) biosensor. We fabricated the GSH exterior on gold surfaces by successive modification with aminoethanethiol, 4‐maleimidobutyric acid N‐hydro...
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| Veröffentlicht in: | Proteomics (Weinheim) 2006-02, Vol.6 (4), p.1110-1120 |
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| creator | Jung, Jae-Wan Jung, Se-Hui Kim, Hyun-Soo Yuk, Jong Seol Park, Jae-Bong Kim, Young-Myeong Han, Jeong-A Kim, Pyung-Hyun Ha, Kwon-Soo |
| description | We modified gold arrays with a glutathione (GSH) surface, and investigated high‐throughput protein interactions with a spectral surface plasmon resonance (SPR) biosensor. We fabricated the GSH exterior on gold surfaces by successive modification with aminoethanethiol, 4‐maleimidobutyric acid N‐hydroxysuccinimide ester and GSH. We immobilized GST‐Rac1, GST‐RhoA, the GST‐Rho‐binding domain of rhotekin and the GST‐p21‐binding domain of PAK1 onto the GSH surface, and observed specific antigen‐antibody interactions on the GST‐fusion protein arrays. We determined the expression of GST‐fusion proteins in Escherichia coli on the GSH surface with the SPR biosensor. We then analyzed the interactions of tissue transglutaminase (tTGase), a Ca2+‐dependent enzyme, with RhoA and Rac1 on the GST‐fusion protein arrays with the SPR biosensor. We found that tTGase interacted with RhoA and Rac1 in a Ca2+‐dependent manner, indicating that the interactions were dependent on tTGase activity. In addition, transamidation of Rac1 by tTGase was dependent on Ca2+ concentration. We obtained similar results with GST pull‐down assays. Thus, protein arrays prepared on the GSH surface provide a useful system for the high‐throughput analysis of GST‐fusion protein expression and activity‐dependent protein interactions with the spectral SPR biosensors. |
| doi_str_mv | 10.1002/pmic.200500371 |
| format | Article |
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We fabricated the GSH exterior on gold surfaces by successive modification with aminoethanethiol, 4‐maleimidobutyric acid N‐hydroxysuccinimide ester and GSH. We immobilized GST‐Rac1, GST‐RhoA, the GST‐Rho‐binding domain of rhotekin and the GST‐p21‐binding domain of PAK1 onto the GSH surface, and observed specific antigen‐antibody interactions on the GST‐fusion protein arrays. We determined the expression of GST‐fusion proteins in Escherichia coli on the GSH surface with the SPR biosensor. We then analyzed the interactions of tissue transglutaminase (tTGase), a Ca2+‐dependent enzyme, with RhoA and Rac1 on the GST‐fusion protein arrays with the SPR biosensor. We found that tTGase interacted with RhoA and Rac1 in a Ca2+‐dependent manner, indicating that the interactions were dependent on tTGase activity. In addition, transamidation of Rac1 by tTGase was dependent on Ca2+ concentration. We obtained similar results with GST pull‐down assays. Thus, protein arrays prepared on the GSH surface provide a useful system for the high‐throughput analysis of GST‐fusion protein expression and activity‐dependent protein interactions with the spectral SPR biosensors.</description><identifier>ISSN: 1615-9853</identifier><identifier>EISSN: 1615-9861</identifier><identifier>DOI: 10.1002/pmic.200500371</identifier><identifier>PMID: 16402361</identifier><language>eng</language><publisher>Weinheim: WILEY-VCH Verlag</publisher><subject>Biosensing Techniques ; Escherichia coli ; Glutathione - chemistry ; Glutathione Transferase - metabolism ; Gold - chemistry ; GSH surface ; GST-fusion protein arrays ; Humans ; Intracellular Signaling Peptides and Proteins - metabolism ; p21-Activated Kinases ; Protein Array Analysis ; Protein Interaction Mapping ; Protein interactions ; Protein-Serine-Threonine Kinases - metabolism ; rac1 GTP-Binding Protein - metabolism ; Recombinant Fusion Proteins - metabolism ; rhoA GTP-Binding Protein - metabolism ; Spectral surface plasmon resonance sensor ; Surface Plasmon Resonance ; Tissue transglutaminase ; Transglutaminases - metabolism</subject><ispartof>Proteomics (Weinheim), 2006-02, Vol.6 (4), p.1110-1120</ispartof><rights>Copyright © 2006 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4121-ffe3e411c63adf5ffc44b55c8ba141c41d424a0caa2448c7151308a89d3c11633</citedby><cites>FETCH-LOGICAL-c4121-ffe3e411c63adf5ffc44b55c8ba141c41d424a0caa2448c7151308a89d3c11633</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fpmic.200500371$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16402361$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jung, Jae-Wan</creatorcontrib><creatorcontrib>Jung, Se-Hui</creatorcontrib><creatorcontrib>Kim, Hyun-Soo</creatorcontrib><creatorcontrib>Yuk, Jong Seol</creatorcontrib><creatorcontrib>Park, Jae-Bong</creatorcontrib><creatorcontrib>Kim, Young-Myeong</creatorcontrib><creatorcontrib>Han, Jeong-A</creatorcontrib><creatorcontrib>Kim, Pyung-Hyun</creatorcontrib><creatorcontrib>Ha, Kwon-Soo</creatorcontrib><title>High-throughput analysis of GST-fusion protein expression and activity-dependent protein interactions on GST-fusion protein arrays with a spectral surface plasmon resonance biosensor</title><title>Proteomics (Weinheim)</title><addtitle>Proteomics</addtitle><description>We modified gold arrays with a glutathione (GSH) surface, and investigated high‐throughput protein interactions with a spectral surface plasmon resonance (SPR) biosensor. We fabricated the GSH exterior on gold surfaces by successive modification with aminoethanethiol, 4‐maleimidobutyric acid N‐hydroxysuccinimide ester and GSH. We immobilized GST‐Rac1, GST‐RhoA, the GST‐Rho‐binding domain of rhotekin and the GST‐p21‐binding domain of PAK1 onto the GSH surface, and observed specific antigen‐antibody interactions on the GST‐fusion protein arrays. We determined the expression of GST‐fusion proteins in Escherichia coli on the GSH surface with the SPR biosensor. We then analyzed the interactions of tissue transglutaminase (tTGase), a Ca2+‐dependent enzyme, with RhoA and Rac1 on the GST‐fusion protein arrays with the SPR biosensor. We found that tTGase interacted with RhoA and Rac1 in a Ca2+‐dependent manner, indicating that the interactions were dependent on tTGase activity. In addition, transamidation of Rac1 by tTGase was dependent on Ca2+ concentration. We obtained similar results with GST pull‐down assays. Thus, protein arrays prepared on the GSH surface provide a useful system for the high‐throughput analysis of GST‐fusion protein expression and activity‐dependent protein interactions with the spectral SPR biosensors.</description><subject>Biosensing Techniques</subject><subject>Escherichia coli</subject><subject>Glutathione - chemistry</subject><subject>Glutathione Transferase - metabolism</subject><subject>Gold - chemistry</subject><subject>GSH surface</subject><subject>GST-fusion protein arrays</subject><subject>Humans</subject><subject>Intracellular Signaling Peptides and Proteins - metabolism</subject><subject>p21-Activated Kinases</subject><subject>Protein Array Analysis</subject><subject>Protein Interaction Mapping</subject><subject>Protein interactions</subject><subject>Protein-Serine-Threonine Kinases - metabolism</subject><subject>rac1 GTP-Binding Protein - metabolism</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>rhoA GTP-Binding Protein - metabolism</subject><subject>Spectral surface plasmon resonance sensor</subject><subject>Surface Plasmon Resonance</subject><subject>Tissue transglutaminase</subject><subject>Transglutaminases - metabolism</subject><issn>1615-9853</issn><issn>1615-9861</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUFv0zAYhi0EYqNw5Yh84pbOX-w4yRFVrJ00BoIBR-urY6-G1Mlshy1_jN9HulaFA9JOtj4_72NbLyGvgc2Bsfys3zo9zxkrGOMlPCGnIKHI6krC0-O-4CfkRYw_GIOyqsvn5ASkYDmXcEp-r9zNJkub0A03m35IFD22Y3SRdpYuv1xndoiu87QPXTLOU3PfBxMfRugbijq5Xy6NWWN64xvj05F0PpmwO-_8JPP_k2EIOEZ659KGIo290SlgS-MQLGpD-xbjdsKnCzuPfpqsXReNj114SZ5ZbKN5dVhn5Ov5--vFKrv8uLxYvLvMtIAcMmsNNwJAS46NLazVQqyLQldrBAET04hcINOIuRCVLqEAziqs6oZrAMn5jLzde6cn3w4mJrV1UZu2RW-6ISpZSglcFo-CUJclKyf9jMz3oA5djMFY1Qe3xTAqYGpXqdpVqo6VToE3B_Ow3prmL37ocALqPXDnWjM-olOfPlws_pVn-6yLydwfsxh-Tl_jZaG-Xy3VisM3Js-v1Gf-B8WHwjE</recordid><startdate>20060201</startdate><enddate>20060201</enddate><creator>Jung, Jae-Wan</creator><creator>Jung, Se-Hui</creator><creator>Kim, Hyun-Soo</creator><creator>Yuk, Jong Seol</creator><creator>Park, Jae-Bong</creator><creator>Kim, Young-Myeong</creator><creator>Han, Jeong-A</creator><creator>Kim, Pyung-Hyun</creator><creator>Ha, Kwon-Soo</creator><general>WILEY-VCH Verlag</general><general>WILEY‐VCH Verlag</general><scope>BSCLL</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>7QL</scope><scope>7QO</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20060201</creationdate><title>High-throughput analysis of GST-fusion protein expression and activity-dependent protein interactions on GST-fusion protein arrays with a spectral surface plasmon resonance biosensor</title><author>Jung, Jae-Wan ; Jung, Se-Hui ; Kim, Hyun-Soo ; Yuk, Jong Seol ; Park, Jae-Bong ; Kim, Young-Myeong ; Han, Jeong-A ; Kim, Pyung-Hyun ; Ha, Kwon-Soo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4121-ffe3e411c63adf5ffc44b55c8ba141c41d424a0caa2448c7151308a89d3c11633</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Biosensing Techniques</topic><topic>Escherichia coli</topic><topic>Glutathione - chemistry</topic><topic>Glutathione Transferase - metabolism</topic><topic>Gold - chemistry</topic><topic>GSH surface</topic><topic>GST-fusion protein arrays</topic><topic>Humans</topic><topic>Intracellular Signaling Peptides and Proteins - metabolism</topic><topic>p21-Activated Kinases</topic><topic>Protein Array Analysis</topic><topic>Protein Interaction Mapping</topic><topic>Protein interactions</topic><topic>Protein-Serine-Threonine Kinases - metabolism</topic><topic>rac1 GTP-Binding Protein - metabolism</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>rhoA GTP-Binding Protein - metabolism</topic><topic>Spectral surface plasmon resonance sensor</topic><topic>Surface Plasmon Resonance</topic><topic>Tissue transglutaminase</topic><topic>Transglutaminases - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Jung, Jae-Wan</creatorcontrib><creatorcontrib>Jung, Se-Hui</creatorcontrib><creatorcontrib>Kim, Hyun-Soo</creatorcontrib><creatorcontrib>Yuk, Jong Seol</creatorcontrib><creatorcontrib>Park, Jae-Bong</creatorcontrib><creatorcontrib>Kim, Young-Myeong</creatorcontrib><creatorcontrib>Han, Jeong-A</creatorcontrib><creatorcontrib>Kim, Pyung-Hyun</creatorcontrib><creatorcontrib>Ha, Kwon-Soo</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Proteomics (Weinheim)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jung, Jae-Wan</au><au>Jung, Se-Hui</au><au>Kim, Hyun-Soo</au><au>Yuk, Jong Seol</au><au>Park, Jae-Bong</au><au>Kim, Young-Myeong</au><au>Han, Jeong-A</au><au>Kim, Pyung-Hyun</au><au>Ha, Kwon-Soo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>High-throughput analysis of GST-fusion protein expression and activity-dependent protein interactions on GST-fusion protein arrays with a spectral surface plasmon resonance biosensor</atitle><jtitle>Proteomics (Weinheim)</jtitle><addtitle>Proteomics</addtitle><date>2006-02-01</date><risdate>2006</risdate><volume>6</volume><issue>4</issue><spage>1110</spage><epage>1120</epage><pages>1110-1120</pages><issn>1615-9853</issn><eissn>1615-9861</eissn><abstract>We modified gold arrays with a glutathione (GSH) surface, and investigated high‐throughput protein interactions with a spectral surface plasmon resonance (SPR) biosensor. We fabricated the GSH exterior on gold surfaces by successive modification with aminoethanethiol, 4‐maleimidobutyric acid N‐hydroxysuccinimide ester and GSH. We immobilized GST‐Rac1, GST‐RhoA, the GST‐Rho‐binding domain of rhotekin and the GST‐p21‐binding domain of PAK1 onto the GSH surface, and observed specific antigen‐antibody interactions on the GST‐fusion protein arrays. We determined the expression of GST‐fusion proteins in Escherichia coli on the GSH surface with the SPR biosensor. We then analyzed the interactions of tissue transglutaminase (tTGase), a Ca2+‐dependent enzyme, with RhoA and Rac1 on the GST‐fusion protein arrays with the SPR biosensor. We found that tTGase interacted with RhoA and Rac1 in a Ca2+‐dependent manner, indicating that the interactions were dependent on tTGase activity. In addition, transamidation of Rac1 by tTGase was dependent on Ca2+ concentration. We obtained similar results with GST pull‐down assays. Thus, protein arrays prepared on the GSH surface provide a useful system for the high‐throughput analysis of GST‐fusion protein expression and activity‐dependent protein interactions with the spectral SPR biosensors.</abstract><cop>Weinheim</cop><pub>WILEY-VCH Verlag</pub><pmid>16402361</pmid><doi>10.1002/pmic.200500371</doi><tpages>11</tpages></addata></record> |
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| ispartof | Proteomics (Weinheim), 2006-02, Vol.6 (4), p.1110-1120 |
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| subjects | Biosensing Techniques Escherichia coli Glutathione - chemistry Glutathione Transferase - metabolism Gold - chemistry GSH surface GST-fusion protein arrays Humans Intracellular Signaling Peptides and Proteins - metabolism p21-Activated Kinases Protein Array Analysis Protein Interaction Mapping Protein interactions Protein-Serine-Threonine Kinases - metabolism rac1 GTP-Binding Protein - metabolism Recombinant Fusion Proteins - metabolism rhoA GTP-Binding Protein - metabolism Spectral surface plasmon resonance sensor Surface Plasmon Resonance Tissue transglutaminase Transglutaminases - metabolism |
| title | High-throughput analysis of GST-fusion protein expression and activity-dependent protein interactions on GST-fusion protein arrays with a spectral surface plasmon resonance biosensor |
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