Polycations globally enhance binding of 14-3-3omega to target proteins in spinach leaves

Plant & Cell Physiology
Wei Shen, S C Huber

Abstract

The binding of 14-3-3omega to phosphorylated nitrate reductase (pNR) is stimulated by cations such as Mg(2+) or spermine, and decreased by 5'-AMP. In order to determine whether binding to other cellular proteins is affected similarly, far-Western overlays of extracts prepared from light- or dark-treated spinach (Spinacia oleracea) leaves were performed using digoxigenin (DIG)-labeled Arabidopsis 14-3-3omega. When separated by SDS-PAGE, approximately 25 proteins of >35 kDa could be resolved that interacted with DIG-labeled 14-3-3omega in the absence of added cations. The presence of 5 mM Mg(2+) or 0.5 mM spermine enhanced binding to most of the target proteins to a maximum of about a doubling of the observed binding. In most cases, the binding was dependent on phosphorylation of the target protein, whereas that was not necessarily the case for binding to target proteins that were unaffected by polycations. The extent of stimulation varied among the target proteins, but there was no indication that the nature of the cation activator (e.g. Mg(2+) vs. spermine(4+)) altered the specificity for target proteins. In addition, binding of DIG-labeled 14-3-3omega to some, but not all, target proteins was reduced by 5 mM 5'-AMP. Interestin...Continue Reading

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Citations

Aug 5, 2011·Amino Acids·Eliana Paola RosalesMaría Patricia Benavides
Jan 26, 2007·Plant & Cell Physiology·Alessandra GarufiPatrizia Aducci
Jan 29, 2014·Chemical Reviews·Russ HillePartha Basu
Jul 7, 2009·Annual Review of Plant Biology·David ChevalierJohn C Walker
Apr 19, 2011·Plant Science : an International Journal of Experimental Plant Biology·Małgorzata RedaGrażyna Kłobus
Apr 26, 2011·Coordination Chemistry Reviews·Russ HilleFlorian Bittner
Sep 13, 2011·Seminars in Cell & Developmental Biology·Fiona C DenisonRobert J Ferl
Jan 16, 2015·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Luisa B Maia, José J G Moura
Apr 4, 2014·Chemical Reviews·Luisa B Maia, José J G Moura
Feb 23, 2018·Frontiers in Chemistry·Avtar K HandaAutar K Mattoo
Nov 14, 2021·Plant Cell Reports·Ye HuangLiang Chen

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