title: A major integral protein of the plant plasma membrane binds flavin.
abstract: Abundant flavin binding sites have been found in membranes of plants and fungi. With flavin mononucleotide-agarose affinity columns, riboflavin-binding activity from microsomes of Cucurbita pepoL. hypocotyls was purified and identified as a specific PIP1-homologous protein of the aquaporin family. Sequences such as gi|2149955 in Phaseolus vulgaris, PIP1b of Arabidopsis thaliana, and NtAQP1 of tobacco are closely related. The identification as a riboflavin-binding protein was confirmed by binding tests with an extract of Escherichia coli cells expressing the tobacco NtAQP1 as well as leaves of transgenic tobacco plants that overexpress NtAQP1 or were inhibited in PIP1 expression by antisense constructs. When binding was assayed in the presence of dithionite, the reduced flavin formed a relatively stable association with the protein. Upon dilution under oxidizing conditions, the adduct was resolved, and free flavin reappeared with a half time of about 30 min. Such an association can also be induced photochemically, with oxidized flavin by blue light at 450 nm, in the presence of an electron donor. Several criteria, localization in the plasma membrane, high abundance, affinity to roseoflavin, and photochemistry, argue for a role of the riboflavin-binding protein PIP1 as a photoreceptor.
authors.affiliation: Institut für Biologie III, Universität Freiburg, Freiburg im Breisgau, Federal Republic of Germany.
authors.forename: Astrid
authors.initials: A
authors.lastname: Lorenz
authors.forename: Ralf
authors.initials: R
authors.lastname: Kaldenhoff
authors.forename: Rainer
authors.initials: R
authors.lastname: Hertel
mesh_terms: D000595:Amino Acid Sequence; D020346:Aquaporins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism; D000440:Butanols; D002462:Cell Membrane / Q000378:metabolism; D002846:Chromatography, Affinity; D004396:Coloring Agents; D028464:Cucurbita / Q000737:chemistry*; D004227:Dithionite; D004983:Ethanolamines; D005486:Flavin Mononucleotide; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D010777:Photochemistry; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D012256:Riboflavin / Q000378:metabolism*; D012685:Sepharose / Q000031:analogs & derivatives*; D020539:Sequence Analysis, Protein; D012995:Solubility; D014316:Tritium; D014357:Trypsin
chemical_list: D020346:Aquaporins; D029681:Arabidopsis Proteins; D000440:Butanols; D004396:Coloring Agents; D004227:Dithionite; D004983:Ethanolamines; D005486:Flavin Mononucleotide; D008565:Membrane Proteins; D010940:Plant Proteins; D012256:Riboflavin; D012685:Sepharose; D014316:Tritium; D014357:Trypsin; C085550:At2g45960 protein, Arabidopsis; C029483:Blue-Sepharose; C007369:2-diethylaminoethanol