dc.creator | Solovou T.G.A., Garagounis C., Kyriakis E., Bobas C., Papadopoulos G.E., Skamnaki V.T., Papadopoulou K.K., Leonidas D.D. | en |
dc.date.accessioned | 2023-01-31T09:58:57Z | |
dc.date.available | 2023-01-31T09:58:57Z | |
dc.date.issued | 2021 | |
dc.identifier | 10.1016/j.phytochem.2021.112707 | |
dc.identifier.issn | 00319422 | |
dc.identifier.uri | http://hdl.handle.net/11615/79189 | |
dc.description.abstract | The glycogen synthase kinases 3 family (GSK3s/SKs; serine/threonine protein kinases) is conserved throughout eukaryotic evolution from yeast to plants and mammals. We studied a plant SK kinase from Lotus japonicus (LjSK1), previously implicated in nodule development, by enzyme kinetics and mutagenesis studies to compare it to mammalian homologues. Using a phosphorylated peptide as substrate, LjSK1 displays optimum kinase activity at pH 8.0 and 20 °C following Michaelis-Menten kinetics with Km and Vmax values of 48.2 μM and 111.6 nmol/min/mg, respectively, for ATP. Mutation of critical residues, as inferred by sequence comparison to the human homologue GSK3β and molecular modeling, showed a conserved role for Lys167, while residues conferring substrate specificity in the human enzyme are not as significant in modulating LjSK1 substrate specificity. Mutagenesis studies also indicate a regulation mechanism for LjSK1 via proteolysis since removal of a 98 residue long N-terminal segment increases its catalytic efficiency by almost two-fold. In addition, we evaluated the alteration of LjSK1 kinase activity in planta, by overexpressing the mutant variants in hairy-roots and a phenotype in nodulation and lateral root development was verified. © 2021 Elsevier Ltd | en |
dc.language.iso | en | en |
dc.source | Phytochemistry | en |
dc.source.uri | https://www.scopus.com/inward/record.uri?eid=2-s2.0-85102316427&doi=10.1016%2fj.phytochem.2021.112707&partnerID=40&md5=797bfaf2c37edfdb4bbaec002232eec0 | |
dc.subject | glycogen synthase kinase 3beta | en |
dc.subject | plant protein | en |
dc.subject | genetics | en |
dc.subject | Lotus (genus) | en |
dc.subject | metabolism | en |
dc.subject | mutagenesis | en |
dc.subject | phosphorylation | en |
dc.subject | Glycogen Synthase Kinase 3 beta | en |
dc.subject | Lotus | en |
dc.subject | Mutagenesis | en |
dc.subject | Phosphorylation | en |
dc.subject | Plant Proteins | en |
dc.subject | Elsevier Ltd | en |
dc.title | Mutagenesis of a Lotus japonicus GSK3β/Shaggy-like kinase reveals functionally conserved regulatory residues | en |
dc.type | journalArticle | en |