A phage display technique for a fast, sensitive, and systematic investigation of protein-protein interactions
Phage display is a technique in which a foreign protein or peptide is presented at the surface of a (filamentous) bacteriophage. This system, developed by Smith [(1985), Science 228, 1315-1317], was originally used to create large libraries of antibodies for the purpose of selecting those that stron...
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| Veröffentlicht in: | Journal of Protein Chemistry 1997-07, Vol.16 (5), p.499-503 |
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| container_title | Journal of Protein Chemistry |
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| creator | Rossenu, S Dewitte, D Vandekerckhove, J Ampe, C |
| description | Phage display is a technique in which a foreign protein or peptide is presented at the surface of a (filamentous) bacteriophage. This system, developed by Smith [(1985), Science 228, 1315-1317], was originally used to create large libraries of antibodies for the purpose of selecting those that strongly bound a particular antigen. More recently it was also employed to present peptides, domains of proteins, or intact proteins at the surface of phages, again to identify high-affinity interactions with ligands. Here we want to illustrate the use of phage display, in combination with PCR saturation mutagenesis, for the study of protein-protein interactions. Rather than selecting for mutants having high affinity, we systematically investigate the binding of every variant with its natural ligand. Via a modified ELISA we can calculate a relative affinity. As a model system we chose to display thymosin beta 4 on the phage surface in order to study its interaction with actin. |
| doi_str_mv | 10.1023/a:1026317612554 |
| format | Article |
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As a model system we chose to display thymosin beta 4 on the phage surface in order to study its interaction with actin.</description><identifier>ISSN: 0277-8033</identifier><identifier>ISSN: 1572-3887</identifier><identifier>EISSN: 1573-4943</identifier><identifier>DOI: 10.1023/a:1026317612554</identifier><identifier>PMID: 9246635</identifier><language>eng</language><publisher>United States: Springer Nature B.V</publisher><subject>Actin ; Actins - metabolism ; Affinity ; Amino Acid Sequence ; Antibodies ; Antigens ; Bacteriophages - genetics ; Bacteriophages - metabolism ; Binding Sites ; Enzyme-Linked Immunosorbent Assay ; Kinetics ; Ligands ; Molecular biology ; Molecular Sequence Data ; Mutagenesis ; Mutation ; Peptides ; Phage display ; Phages ; Polymerase Chain Reaction ; Protein interaction ; Proteins ; Proteins - genetics ; Proteins - metabolism ; Saturation mutagenesis ; Sensitivity and Specificity ; Thymosin - genetics ; Thymosin - metabolism</subject><ispartof>Journal of Protein Chemistry, 1997-07, Vol.16 (5), p.499-503</ispartof><rights>Plenum Publishing Corporation 1997.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c488t-5e23aa35d0b64f04c6ad49ca7d8e86916d3ea7611887c8241e8324bd50150fad3</citedby><cites>FETCH-LOGICAL-c488t-5e23aa35d0b64f04c6ad49ca7d8e86916d3ea7611887c8241e8324bd50150fad3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9246635$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rossenu, S</creatorcontrib><creatorcontrib>Dewitte, D</creatorcontrib><creatorcontrib>Vandekerckhove, J</creatorcontrib><creatorcontrib>Ampe, C</creatorcontrib><title>A phage display technique for a fast, sensitive, and systematic investigation of protein-protein interactions</title><title>Journal of Protein Chemistry</title><addtitle>J Protein Chem</addtitle><description>Phage display is a technique in which a foreign protein or peptide is presented at the surface of a (filamentous) bacteriophage. This system, developed by Smith [(1985), Science 228, 1315-1317], was originally used to create large libraries of antibodies for the purpose of selecting those that strongly bound a particular antigen. More recently it was also employed to present peptides, domains of proteins, or intact proteins at the surface of phages, again to identify high-affinity interactions with ligands. Here we want to illustrate the use of phage display, in combination with PCR saturation mutagenesis, for the study of protein-protein interactions. Rather than selecting for mutants having high affinity, we systematically investigate the binding of every variant with its natural ligand. Via a modified ELISA we can calculate a relative affinity. As a model system we chose to display thymosin beta 4 on the phage surface in order to study its interaction with actin.</description><subject>Actin</subject><subject>Actins - metabolism</subject><subject>Affinity</subject><subject>Amino Acid Sequence</subject><subject>Antibodies</subject><subject>Antigens</subject><subject>Bacteriophages - genetics</subject><subject>Bacteriophages - metabolism</subject><subject>Binding Sites</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Kinetics</subject><subject>Ligands</subject><subject>Molecular biology</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis</subject><subject>Mutation</subject><subject>Peptides</subject><subject>Phage display</subject><subject>Phages</subject><subject>Polymerase Chain Reaction</subject><subject>Protein interaction</subject><subject>Proteins</subject><subject>Proteins - genetics</subject><subject>Proteins - metabolism</subject><subject>Saturation mutagenesis</subject><subject>Sensitivity and Specificity</subject><subject>Thymosin - genetics</subject><subject>Thymosin - 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| identifier | ISSN: 0277-8033 |
| ispartof | Journal of Protein Chemistry, 1997-07, Vol.16 (5), p.499-503 |
| issn | 0277-8033 1572-3887 1573-4943 |
| language | eng |
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| source | MEDLINE; SpringerLink Journals - AutoHoldings |
| subjects | Actin Actins - metabolism Affinity Amino Acid Sequence Antibodies Antigens Bacteriophages - genetics Bacteriophages - metabolism Binding Sites Enzyme-Linked Immunosorbent Assay Kinetics Ligands Molecular biology Molecular Sequence Data Mutagenesis Mutation Peptides Phage display Phages Polymerase Chain Reaction Protein interaction Proteins Proteins - genetics Proteins - metabolism Saturation mutagenesis Sensitivity and Specificity Thymosin - genetics Thymosin - metabolism |
| title | A phage display technique for a fast, sensitive, and systematic investigation of protein-protein interactions |
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