title: Molecular characterization of a FKBP-type immunophilin from higher plants.
abstract: Immunophilins are intracellular receptors for the immunosuppressants cyclosporin A, FK506, and rapamycin. In addition to their use in organ transplantation, these natural products have been used to investigate signaling pathways in yeast, plant, and mammalian cells. We have recently described the identification of an immunosuppressant-sensitive signaling pathway in and the purification of several immunophilins from Vicia faba plants. We now report the molecular characterization of a 15 kDa FK506- and rapamycin-binding protein from V. faba (VfFKBP15). The amino acid sequence deduced from the cDNA starts with a signal peptide of 22 hydrophobic amino acids. The core region of VfFKBP15 is most similar to yeast and mammalian FKBP13 localized in the endoplasmic reticulum (ER). In addition, VfFKBP15 has a carboxyl-terminal sequence that is ended with SSEL, a putative ER retention signal. These findings suggest that VfFKBP15 is a functional homolog of FKBP13 from other organisms. Interestingly, two distinct cDNAs corresponding to two isoforms of FKBP15 have been cloned from Arabidopsis and also identified from rice data base, suggesting that pFKBP15 (plant FKBP15) is encoded by a small gene family in plants. This adds to the diversity of plant FKBP members even with the same subcellular localization and is in contrast with the situation in mammalian and yeast systems in which only one FKBP13 gene has been found. Like the mammalian and yeast FKBP13, the recombinant VfFKBP15 protein has rotamase activity that is inhibited by both FK506 and rapamycin with a Ki value of 30 nM and 0.9 nM, respectively, illustrating that VfFKBP15 binds rapamycin in preference over FK506. The mRNA of VfFKBP15 is ubiquitously expressed in various plant tissues including leaves, stems, and roots, consistent with the ER localization of the protein. Levels of VfFKBP15 mRNA are elevated by heat shock, suggesting a possible role for this FKBP member under stress conditions.
authors.affiliation: Department of Plant Biology, University of California, Berkeley, CA 94720, USA.
authors.forename: S
authors.initials: S
authors.lastname: Luan
authors.forename: J
authors.initials: J
authors.lastname: Kudla
authors.forename: W
authors.initials: W
authors.lastname: Gruissem
authors.forename: S L
authors.initials: SL
authors.lastname: Schreiber
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D018076:DNA, Complementary; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D007887:Fabaceae / Q000235:genetics / Q000378:metabolism*; D015723:Gene Library; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007166:Immunosuppressive Agents / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D007535:Isomerases / Q000378:metabolism*; D007700:Kinetics; D008322:Mammals; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D018515:Plant Leaves; D010946:Plants, Medicinal; D011090:Polyenes / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae; D017386:Sequence Homology, Amino Acid; D020123:Sirolimus; D013379:Substrate Specificity; D016559:Tacrolimus / Q000378:metabolism; D022021:Tacrolimus Binding Proteins; D014158:Transcription, Genetic
chemical_list: D002352:Carrier Proteins; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D006360:Heat-Shock Proteins; D007166:Immunosuppressive Agents; D015345:Oligonucleotide Probes; D011090:Polyenes; D012333:RNA, Messenger; D007535:Isomerases; D022021:Tacrolimus Binding Proteins; D020123:Sirolimus; D016559:Tacrolimus