{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T07:01:32Z","timestamp":1789714892861,"version":"4.0.1"},"reference-count":34,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2002,1,18]],"date-time":"2002-01-18T00:00:00Z","timestamp":1011312000000},"content-version":"vor","delay-in-days":688,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Microbiology"],"published-print":{"date-parts":[[2000,3]]},"abstract":"<jats:p>The commercially important bacterium <jats:italic>Lactococcus lactis<\/jats:italic> contains two FNR\u2010like proteins (FlpA and FlpB) which have a high degree of identity to each other and to the FLP of <jats:italic>Lactobacillus casei<\/jats:italic>. FlpA was isolated from a GST\u2013FlpA fusion protein produced in <jats:italic>Escherichia coli<\/jats:italic>. Like FLP, isolated FlpA is a homodimeric protein containing both Zn and Cu. However, the properties of FlpA were more like those of the <jats:italic>E. coli<\/jats:italic> oxygen\u2010responsive transcription factor FNR than the FLP of <jats:italic>L. casei<\/jats:italic>. As prepared FlpA recognized an FNR site (TTGAT\u2010N4\u2010ATCAA) but not an FLP site (CCTGA\u2010N4\u2010TCAGG) in band\u2010shift assays. In contrast to FLP, DNA binding by FlpA did not require the formation of an intramolecular disulphide bond. However, despite containing only two cysteine residues per monomer, FlpA was able to acquire an FNR\u2010like, oxygen\u2010labile [4Fe 4S] cluster. But, whereas the incorporation of a [4Fe 4S] cluster into FNR enhances interaction with target DNA, it abolished DNA binding by FlpA. An FlpA variant (FlpA\u2032) with an N\u2010terminal region designed to be more FLP\u2010like failed to incorporate an iron\u2013sulphur cluster but could now form an intramolecular disulphide. This simple example of protein engineering, converting an oxygen\u2010labile [4Fe 4S] containing FNR\u2010like protein into a dithiol\u2013disulphide FLP\u2010like redox sensor demonstrates the versatility of the basic CRP structure. Attempts to demonstrate an FlpA\u2010based aerobic\u2013anaerobic switch in the heterologous host <jats:italic>E. coli<\/jats:italic> were unsuccessful. However, studies with a series of FNR\u2010dependent lac reporter fusions in strains of <jats:italic>E. coli<\/jats:italic> expressing <jats:italic>flpA<\/jats:italic> or <jats:italic>flpB<\/jats:italic> revealed that both homologues were able to activate expression of FNR\u2010dependent promoters <jats:italic>in vivo<\/jats:italic> but only when positioned 61 base pairs upstream of the transcription start.<\/jats:p>","DOI":"10.1046\/j.1365-2958.2000.01799.x","type":"journal-article","created":{"date-parts":[[2003,3,12]],"date-time":"2003-03-12T11:29:44Z","timestamp":1047468584000},"page":"1383-1393","source":"Crossref","is-referenced-by-count":27,"title":["Characterization of the <i>Lactococcus lactis<\/i> transcription factor FlpA and demonstration of an <i>in vitro<\/i> switch"],"prefix":"10.1111","volume":"35","author":[{"given":"Colin","family":"Scott","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John R.","family":"Guest","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeffrey","family":"Green","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2002,1,18]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)80584-X"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(83)90186-0"},{"key":"e_1_2_6_4_1","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1016\/S0021-9258(17)43630-1","article-title":"Purification and characterization of the Rieske iron\u2010sulfur protein from Thermus thermophilus","volume":"259","author":"Fee J.A.","year":"1984","journal-title":"J Biol Chem"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6976.1993.tb00010.x"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1099\/00221287-144-3-705"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1999.01298.x"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1997.3801737.x"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1998.01002.x"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj3160887"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.1993.tb01203.x"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1098\/rspb.1991.0062"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/21.3.753"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)80547-4"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(97)01219-2"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.49.31135"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.7.2499"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.173.1.16-22.1991"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/227680a0"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/0042-6822(55)90016-7"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.1990.tb13867.x"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/14.8.3603"},{"key":"e_1_2_6_23_1","volume-title":"Experiments in Molecular Genetics.","author":"Miller J.H. 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