title: Birch PR-10c interacts with several biologically important ligands.
abstract: PR-10c is a unique member of PR-10 proteins in birch, since it is the only one known to be post-translationally modified by glutathione and is not constitutively expressed in pollen. Both reduced and S-glutathiolated forms of PR-10c show low ribonuclease activity. However, the major function of the protein is apparently not yet resolved. Our protein-ligand interaction studies with saturation transfer difference (STD) NMR revealed that PR-10c interacts with several biologically important molecules, including cytokinin, flavonoid glycosides, sterols and emodin. Competition study with deoxycholate and kinetin revealed no statistically significant binding interference, indicating that these ligands have different binding sites in PR-10c. Ligand docking studies with a molecular model of PR-10c support the STD NMR results of ligand binding and binding epitopes, suggesting that there are three potential binding sites in PR-10c: two in the hydrophobic cavity and one in the glycine-rich loop. Our docking calculations suggested that only kinetin interacts with the glycine-rich loop, the binding occurring through its adenine moiety. Clear ligand specificity could be observed in the binding of nucleotide derivatives. S-glutathiolation of PR-10c did not affect kinetin binding. The present results suggest that birch PR-10c is a multifunctional protein, which has diverse roles in plant stress responses.
authors.affiliation: Institute of Applied Biotechnology, University of Kuopio, P.O. Box 1627, FIN-70211 Kuopio, Finland.
authors.forename: Kaisa M
authors.initials: KM
authors.lastname: Koistinen
authors.forename: Pasi
authors.initials: P
authors.lastname: Soininen
authors.forename: Tuomas A
authors.initials: TA
authors.lastname: Venäläinen
authors.forename: Jukka
authors.initials: J
authors.lastname: Häyrinen
authors.forename: Reino
authors.initials: R
authors.lastname: Laatikainen
authors.forename: Mikael
authors.initials: M
authors.lastname: Peräkylä
authors.forename: Arja I
authors.initials: AI
authors.lastname: Tervahauta
authors.forename: Sirpa O
authors.initials: SO
authors.lastname: Kärenlampi
mesh_terms: D029662:Betula; D003840:Deoxycholic Acid / Q000378:metabolism; D004642:Emodin / Q000378:metabolism; D007701:Kinetin / Q000378:metabolism; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D010940:Plant Proteins / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D011794:Quercetin / Q000031:analogs & derivatives / Q000378:metabolism; D012431:Rutin / Q000378:metabolism
chemical_list: D008024:Ligands; D010940:Plant Proteins; D003840:Deoxycholic Acid; D012431:Rutin; C021304:hyperoside; D011794:Quercetin; D004642:Emodin; D007701:Kinetin