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The platform is based on a versatile 42\u2010residue helix\u2013loop\u2013helix polypeptide that dimerizes to form four\u2010helix bundles and allows site\u2010selective modification with recognition and reporter elements on the side chains of individually addressable lysine residues. The well\u2010characterized interaction between the model target\u2010protein carbonic anhydrase and its inhibitor benzenesulfonamide was used for a proof\u2010of\u2010concept demonstration. An affinity array was designed where benzenesulfonamide derivatives with aliphatic or oligoglycine spacers and a fluorescent dansyl reporter group were introduced into the scaffold. The affinities of the array members for human carbonic anhydrase II (HCAII) were determined by titration with the target protein and were found to be highly affected by the properties of the spacers (dissociation constant <jats:italic>K<\/jats:italic><jats:sub>d<\/jats:sub>=0.02\u20133 \u03bc<jats:sc>M<\/jats:sc>). The affinity of HCAII for acetazolamide (<jats:italic>K<\/jats:italic><jats:sub>d<\/jats:sub>=4 n<jats:sc>M<\/jats:sc>) was determined in a competition experiment with one of the benzenesulfonamide array members to address the possibility of screening substance libraries for new target\u2010protein binders. Also, successful affinity discrimination between different carbonic anhydrase isozymes highlighted the possibility of performing future isoform\u2010expression profiling. Our platform is predicted to become a flexible tool for a variety of biosensor and protein\u2010microarray applications within biochemistry, diagnostics and pharmaceutical chemistry.<\/jats:p>","DOI":"10.1002\/chem.200305391","type":"journal-article","created":{"date-parts":[[2004,4,15]],"date-time":"2004-04-15T12:06:57Z","timestamp":1082030817000},"page":"2375-2385","source":"Crossref","is-referenced-by-count":29,"title":["A Versatile Polypeptide Platform for Integrated Recognition and Reporting: Affinity Arrays for Protein\u2013Ligand Interaction Analysis"],"prefix":"10.1002","volume":"10","author":[{"given":"Karin","family":"Enander","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gunnar T.","family":"Dolphin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bo","family":"Liedberg","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ingemar","family":"Lundstr\u00f6m","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lars","family":"Baltzer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2004,5,7]]},"reference":[{"key":"e_1_2_7_1_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-5521(00)90067-X"},{"key":"e_1_2_7_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-7799(01)01910-2"},{"key":"e_1_2_7_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1367-5931(02)00005-4"},{"key":"e_1_2_7_4_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi0023201"},{"key":"e_1_2_7_5_2","doi-asserted-by":"publisher","DOI":"10.1002\/1521-3757(20010302)113:5<922::AID-ANGE922>3.0.CO;2-K"},{"key":"e_1_2_7_5_3","doi-asserted-by":"publisher","DOI":"10.1002\/1521-3773(20010302)40:5<897::AID-ANIE897>3.0.CO;2-X"},{"key":"e_1_2_7_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0302-270"},{"key":"e_1_2_7_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-5521(02)00308-3"},{"key":"e_1_2_7_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-5521(03)00049-8"},{"key":"e_1_2_7_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/74442"},{"key":"e_1_2_7_10_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja961097l"},{"key":"e_1_2_7_11_2","doi-asserted-by":"publisher","DOI":"10.1002\/1439-7633(20020802)3:8<741::AID-CBIC741>3.0.CO;2-O"},{"key":"e_1_2_7_12_2","doi-asserted-by":"publisher","DOI":"10.1002\/1521-3765(20020816)8:16<3687::AID-CHEM3687>3.0.CO;2-8"},{"key":"e_1_2_7_13_2","doi-asserted-by":"publisher","DOI":"10.1021\/jo971999k"},{"key":"e_1_2_7_14_2","doi-asserted-by":"publisher","DOI":"10.1039\/B302847J"},{"key":"e_1_2_7_15_2","doi-asserted-by":"publisher","DOI":"10.1021\/jo010954n"},{"key":"e_1_2_7_16_2","doi-asserted-by":"publisher","DOI":"10.1351\/pac199971122333"},{"key":"e_1_2_7_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-7799(98)01174-3"},{"key":"e_1_2_7_18_2","doi-asserted-by":"publisher","DOI":"10.1039\/P29950002047"},{"key":"e_1_2_7_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1359-0278(96)00050-8"},{"key":"e_1_2_7_20_2","unstructured":"L. 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