On the determination of deoxyribonucleic acid-protein interaction parameters using the nitrocellulose filter-binding assay
We examine the effects of filter efficiency on DNA-protein binding data obtained by the popular nitrocellulose filter-binding assay. Graphical procedures for determining the efficiency parameter epsilon (for the efficiency of retention of DNA on the filter, per bound protein molecule) are establishe...
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| Veröffentlicht in: | Biochemistry (Easton) 1983-09, Vol.22 (20), p.4730-4737 |
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| container_end_page | 4737 |
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| container_issue | 20 |
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| container_title | Biochemistry (Easton) |
| container_volume | 22 |
| creator | Woodbury, Charles P Von Hippel, Peter H |
| description | We examine the effects of filter efficiency on DNA-protein binding data obtained by the popular nitrocellulose filter-binding assay. Graphical procedures for determining the efficiency parameter epsilon (for the efficiency of retention of DNA on the filter, per bound protein molecule) are established. Filter efficiency modified formulas for determining thermodynamic binding parameters are derived for four simple prototypes of DNA-protein binding systems. The effects of experimental error on discrimination between models are considered. Finally, we discuss conceptual errors often made in calculating binding stoichiometry from filter-binding experiments and suggest a general protocol for the analysis of filter-binding data. |
| doi_str_mv | 10.1021/bi00289a018 |
| format | Article |
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Graphical procedures for determining the efficiency parameter epsilon (for the efficiency of retention of DNA on the filter, per bound protein molecule) are established. Filter efficiency modified formulas for determining thermodynamic binding parameters are derived for four simple prototypes of DNA-protein binding systems. The effects of experimental error on discrimination between models are considered. Finally, we discuss conceptual errors often made in calculating binding stoichiometry from filter-binding experiments and suggest a general protocol for the analysis of filter-binding data.</description><identifier>ISSN: 0006-2960</identifier><identifier>EISSN: 1520-4995</identifier><identifier>DOI: 10.1021/bi00289a018</identifier><identifier>PMID: 6626528</identifier><language>eng</language><publisher>Washington, DC: American Chemical Society</publisher><subject>Biological and medical sciences ; DNA - metabolism ; Fundamental and applied biological sciences. 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Graphical procedures for determining the efficiency parameter epsilon (for the efficiency of retention of DNA on the filter, per bound protein molecule) are established. Filter efficiency modified formulas for determining thermodynamic binding parameters are derived for four simple prototypes of DNA-protein binding systems. The effects of experimental error on discrimination between models are considered. Finally, we discuss conceptual errors often made in calculating binding stoichiometry from filter-binding experiments and suggest a general protocol for the analysis of filter-binding data.</description><subject>Biological and medical sciences</subject><subject>DNA - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Interactions. Associations</subject><subject>Intermolecular phenomena</subject><subject>Kinetics</subject><subject>Mathematics</subject><subject>Models, Biological</subject><subject>Molecular biophysics</subject><subject>Protein Binding</subject><subject>Proteins - metabolism</subject><issn>0006-2960</issn><issn>1520-4995</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1983</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpt0M9rFDEUB_AgSt1WT56FOYg9yOhLZiaTHGvRVihWaQVv4U1-aOpMsiYz0PWvN9tdFg-eQvL95PH4EvKCwlsKjL4bPAATEoGKR2RFOwZ1K2X3mKwAgNdMcnhKjnO-K9cW-vaIHHHOeMfEivy5DtX801bGzjZNPuDsY6iiKw_xfpP8EMOiR-t1hdqbep3ibH2ofCgc9QNeY8Jp-z1XS_bhx8O84OcUtR3HZYzZVs6PBdSDD2YrMGfcPCNPHI7ZPt-fJ-Tbxw-355f11fXFp_Ozqxob0cw1c9YywVsU0LkWBofMsA5hMIPErueaciMN5cIw3kvmoNVcINO0l7YHBs0Jeb2bW5b_vdg8q8nn7WoYbFyyEsCl6Bkv8M0O6hRzTtapdfITpo2ioLZNq3-aLvrlfuwyTNYc7L7akr_a55g1ji5h0D4fmGwEa7uusHrHfJ7t_SHG9Evxvuk7dfvlRrGv39_fyIvP6rL4051HndVdXFIo3f13wb-E3qQD</recordid><startdate>19830927</startdate><enddate>19830927</enddate><creator>Woodbury, Charles P</creator><creator>Von Hippel, Peter H</creator><general>American Chemical Society</general><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></search><sort><creationdate>19830927</creationdate><title>On the determination of deoxyribonucleic acid-protein interaction parameters using the nitrocellulose filter-binding assay</title><author>Woodbury, Charles P ; Von Hippel, Peter H</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a383t-2fee2864a805f40bfa2d25a0bdb9a576c16d9d168d26792f04c68a2c179e70203</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1983</creationdate><topic>Biological and medical sciences</topic><topic>DNA - metabolism</topic><topic>Fundamental and applied biological sciences. 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| ispartof | Biochemistry (Easton), 1983-09, Vol.22 (20), p.4730-4737 |
| issn | 0006-2960 1520-4995 |
| language | eng |
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| source | MEDLINE; American Chemical Society Journals |
| subjects | Biological and medical sciences DNA - metabolism Fundamental and applied biological sciences. Psychology Interactions. Associations Intermolecular phenomena Kinetics Mathematics Models, Biological Molecular biophysics Protein Binding Proteins - metabolism |
| title | On the determination of deoxyribonucleic acid-protein interaction parameters using the nitrocellulose filter-binding assay |
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