Dual binding of 14-3-3 protein regulates Arabidopsis nitrate reductase activity
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| 001 | 605507929 | ||
| 003 | CHVBK | ||
| 005 | 20210128100634.0 | ||
| 007 | cr unu---uuuuu | ||
| 008 | 210128e20150301xx s 000 0 eng | ||
| 024 | 7 | 0 | |a 10.1007/s00775-014-1232-4 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00775-014-1232-4 | ||
| 245 | 0 | 0 | |a Dual binding of 14-3-3 protein regulates Arabidopsis nitrate reductase activity |h [Elektronische Daten] |c [Jen-Chih Chi, Juliane Roeper, Guenter Schwarz, Katrin Fischer-Schrader] |
| 520 | 3 | |a 14-3-3 proteins represent a family of ubiquitous eukaryotic proteins involved in numerous signal transduction processes and metabolic pathways. One important 14-3-3 target in higher plants is nitrate reductase (NR), whose activity is regulated by different physiological conditions. Intra-molecular electron transfer in NR is inhibited following 14-3-3 binding to a conserved phospho-serine motif located in hinge 1, a surface exposed loop between the catalytic molybdenum and central heme domain. Here we describe a novel 14-3-3 binding site within the NR N-terminus, an acidic motif conserved in NRs of higher plants, which significantly contributes to 14-3-3-mediated inhibition of NR. Deletion or mutation of the N-terminal acidic motif resulted in a significant loss of 14-3-3 mediated inhibition of Ser534 phosphorylated NR-Mo-heme (residues 1-625), a previously established model of NR regulation. Co-sedimentation and crosslinking studies with NR peptides comprising each of the two binding motifs demonstrated direct binding of either peptide to 14-3-3. Surface plasmon resonance spectroscopy disclosed high-affinity binding of 14-3-3ω to the well-known phospho-hinge site and low-affinity binding to the N-terminal acidic motif. A binding groove-deficient 14-3-3ω variant retained interaction to the acidic motif, but lost binding to the phospho-hinge motif. To our knowledge, NR is the first enzyme that harbors two independent 14-3-3 binding sites with different affinities, which both need to be occupied by 14-3-3ω to confer full inhibition of NR activity under physiological conditions. | |
| 540 | |a SBIC, 2015 | ||
| 690 | 7 | |a Phosphorylation |2 nationallicence | |
| 690 | 7 | |a Acidic motif |2 nationallicence | |
| 690 | 7 | |a 14-3-3 |2 nationallicence | |
| 690 | 7 | |a Nitrate reductase |2 nationallicence | |
| 690 | 7 | |a Regulation |2 nationallicence | |
| 690 | 7 | |a EDC : 1-Ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride |2 nationallicence | |
| 690 | 7 | |a GST : Glutathione-S-transferase |2 nationallicence | |
| 690 | 7 | |a NR : Nitrate reductase |2 nationallicence | |
| 690 | 7 | |a NT : N-terminus of nitrate reductase |2 nationallicence | |
| 690 | 7 | |a SPR : Surface plasmon resonance spectroscopy |2 nationallicence | |
| 690 | 7 | |a wt : Wild-type |2 nationallicence | |
| 700 | 1 | |a Chi |D Jen-Chih |u Department of Chemistry, Institute for Biochemistry, University of Cologne, 50674, Cologne, Germany |4 aut | |
| 700 | 1 | |a Roeper |D Juliane |u Department of Chemistry, Institute for Biochemistry, University of Cologne, 50674, Cologne, Germany |4 aut | |
| 700 | 1 | |a Schwarz |D Guenter |u Department of Chemistry, Institute for Biochemistry, University of Cologne, 50674, Cologne, Germany |4 aut | |
| 700 | 1 | |a Fischer-Schrader |D Katrin |u Department of Chemistry, Institute for Biochemistry, University of Cologne, 50674, Cologne, Germany |4 aut | |
| 773 | 0 | |t JBIC Journal of Biological Inorganic Chemistry |d Springer Berlin Heidelberg |g 20/2(2015-03-01), 277-286 |x 0949-8257 |q 20:2<277 |1 2015 |2 20 |o 775 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00775-014-1232-4 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00775-014-1232-4 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Chi |D Jen-Chih |u Department of Chemistry, Institute for Biochemistry, University of Cologne, 50674, Cologne, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Roeper |D Juliane |u Department of Chemistry, Institute for Biochemistry, University of Cologne, 50674, Cologne, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Schwarz |D Guenter |u Department of Chemistry, Institute for Biochemistry, University of Cologne, 50674, Cologne, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Fischer-Schrader |D Katrin |u Department of Chemistry, Institute for Biochemistry, University of Cologne, 50674, Cologne, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t JBIC Journal of Biological Inorganic Chemistry |d Springer Berlin Heidelberg |g 20/2(2015-03-01), 277-286 |x 0949-8257 |q 20:2<277 |1 2015 |2 20 |o 775 | ||