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Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

The labile zinc pool in plant cells

Ilya E. Zlobin https://orcid.org/0000-0001-9317-9414 A B , Alexander V. Kartashov A , Alexander V. Nosov A , Artem A. Fomenkov A and Vladimir V. Kuznetsov A
+ Author Affiliations
- Author Affiliations

A Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, 127276 Moscow, Russia.

B Corresponding author. Email: ilya.zlobin.90@mail.ru

Functional Plant Biology 46(9) 796-805 https://doi.org/10.1071/FP19064
Submitted: 26 February 2019  Accepted: 18 April 2019   Published: 10 May 2019

Abstract

Zinc is the most abundant and important transition metal in plants; however, the dynamic aspects of zinc homeostasis in plant cells are poorly understood. In this study we explored the pool of labile exchangeable zinc complexes in plant cells, and the potential influence of changes in intracellular zinc availability on cellular physiology. Work was performed on cultivated cell extracts of Arabidopsis thaliana (L.) Heynh. and Thellungiella salsuginea (Pall.) O.E. Schulz grown under control (3.48 µM Zn2+), 10-fold Zn excess or Zn starvation conditions. The free and labile Zn contents in the extracts were then determined by fluorimetric titration. We observed for the first time that plant cells contain micromolar concentrations of labile zinc complexes that account for a low percentage of the total zinc content. Labile zinc is mainly protein bound. Zn starvation inhibits cell proliferation and leads to the disappearance of the labile zinc pool, whereas Zn excess drastically increases the labile zinc pool. Free Zn2+ is buffered at picomolar concentrations in the intracellular milieu, and the increase in free Zn2+ concentrations to low nanomolar values clearly modulates enzyme activity by direct reversible binding. Such increases in free Zn2+ can be achieved by the substantial influx of additional zinc or by the oxidation of zinc-binding thiols. The observed features of the labile zinc pool in plant cells suggest it has a role in intracellular zinc trafficking and zinc signalling.

Additional keywords: enzyme activity, phosphatase, plant signalling, trace metals, zinc metabolism.


References

Adam S, Murthy SDS (2014) Characterization of alterations in photosynthetic electron transport activities in maize thylakoid membranes under zinc stress. European Journal of Experimental Biology 4, 25–29.

Andreini C, Banci L, Bertini I, Rosato A (2006) Counting the zinc-proteins encoded in the human genome. Journal of Proteome Research 5, 196–201.
Counting the zinc-proteins encoded in the human genome.Crossref | GoogleScholarGoogle Scholar | 16396512PubMed |

Blindauer CA (2015) Advances in the molecular understanding of biological zinc transport. Chemical Communications 51, 4544–4563.
Advances in the molecular understanding of biological zinc transport.Crossref | GoogleScholarGoogle Scholar | 25627157PubMed |

Blindauer CA, Schmid R (2010) Cytosolic metal handling in plants: determinants for zinc specificity in metal transporters and metallothioneins. Metallomics 2, 510–529.
Cytosolic metal handling in plants: determinants for zinc specificity in metal transporters and metallothioneins.Crossref | GoogleScholarGoogle Scholar | 21072336PubMed |

Colvin RA, Holmes WR, Fontaine CP, Maret W (2010) Cytosolic zinc buffering and muffling: their role in intracellular zinc homeostasis. Metallomics 2, 306–317.
Cytosolic zinc buffering and muffling: their role in intracellular zinc homeostasis.Crossref | GoogleScholarGoogle Scholar | 21069178PubMed |

Costello LC, Fenselau CC, Franklin RB (2011) Evidence for operation of the direct zinc ligand exchange mechanism for trafficking, transport, and reactivity of zinc in mammalian cells. Journal of Inorganic Biochemistry 105, 589–599.
Evidence for operation of the direct zinc ligand exchange mechanism for trafficking, transport, and reactivity of zinc in mammalian cells.Crossref | GoogleScholarGoogle Scholar | 21440525PubMed |

Deinlein U, Weber M, Schmidt H, Rensch S, Trampczynska A, Hansen TH, Husted S, Schjoerring JK, Talke IN, Krämer U, Clemens S (2012) Elevated nicotianamine levels in Arabidopsis halleri roots play a key role in zinc hyperaccumulation. The Plant Cell 24, 708–723.
Elevated nicotianamine levels in Arabidopsis halleri roots play a key role in zinc hyperaccumulation.Crossref | GoogleScholarGoogle Scholar | 22374395PubMed |

Haase H, Rink L (2009) Functional significance of zinc-related signaling pathways in immune cells. Annual Review of Nutrition 29, 133–152.
Functional significance of zinc-related signaling pathways in immune cells.Crossref | GoogleScholarGoogle Scholar | 19400701PubMed |

Haase H, Hebel S, Engelhardt G, Rink L (2015) The biochemical effects of extracellular Zn2+ and other metal ions are severely affected by their speciation in cell culture media. Metallomics 7, 102–111.
The biochemical effects of extracellular Zn2+ and other metal ions are severely affected by their speciation in cell culture media.Crossref | GoogleScholarGoogle Scholar | 25360687PubMed |

Hara M, Shinoda Y, Tanaka Y, Kuboi T (2009) DNA binding of citrus dehydrin promoted by zinc ion. Plant, Cell & Environment 32, 532–541.
DNA binding of citrus dehydrin promoted by zinc ion.Crossref | GoogleScholarGoogle Scholar |

Heinz U, Kiefer M, Tholey A, Adolph HW (2005) On the competition for available zinc. The Journal of Biological Chemistry 280, 3197–3207.
On the competition for available zinc.Crossref | GoogleScholarGoogle Scholar | 15536071PubMed |

Helmersson A, von Arnold S, Bozhkov PV (2008) The level of free intracellular zinc mediates programmed cell death/cell survival decisions in plant embryos. Plant Physiology 147, 1158–1167.
The level of free intracellular zinc mediates programmed cell death/cell survival decisions in plant embryos.Crossref | GoogleScholarGoogle Scholar | 18508953PubMed |

Kochańczyk T, Drozd A, Krężel A (2015) Relationship between the architecture of zinc coordination and zinc binding affinity in proteins – insights into zinc regulation. Metallomics 7, 244–257.
Relationship between the architecture of zinc coordination and zinc binding affinity in proteins – insights into zinc regulation.Crossref | GoogleScholarGoogle Scholar | 25255078PubMed |

Krężel A, Maret W (2006) Zinc-buffering capacity of a eukaryotic cell at physiological pZn. JBIC Journal of Biological Inorganic Chemistry 11, 1049–1062.
Zinc-buffering capacity of a eukaryotic cell at physiological pZn.Crossref | GoogleScholarGoogle Scholar | 16924557PubMed |

Krężel A, Maret W (2007) Dual nanomolar and picomolar Zn (II) binding properties of metallothionein. Journal of the American Chemical Society 129, 10911–10921.
Dual nanomolar and picomolar Zn (II) binding properties of metallothionein.Crossref | GoogleScholarGoogle Scholar | 17696343PubMed |

Krężel A, Maret W (2016) The biological inorganic chemistry of zinc ions. Archives of Biochemistry and Biophysics 611, 3–19.
The biological inorganic chemistry of zinc ions.Crossref | GoogleScholarGoogle Scholar | 27117234PubMed |

Kühnlenz T, Hofmann C, Uraguchi S, Schmidt H, Schempp S, Weber M, Lahner B, Salt DE, Clemens S (2016) Phytochelatin synthesis promotes leaf Zn accumulation of Arabidopsis thaliana plants grown in soil with adequate Zn supply and is essential for survival on Zn-contaminated soil. Plant & Cell Physiology 57, 2342–2352.
Phytochelatin synthesis promotes leaf Zn accumulation of Arabidopsis thaliana plants grown in soil with adequate Zn supply and is essential for survival on Zn-contaminated soil.Crossref | GoogleScholarGoogle Scholar |

Lanquar V, Grossmann G, Vinkenborg JL, Merkx M, Thomine S, Frommer WB (2014) Dynamic imaging of cytosolic zinc in Arabidopsis roots combining FRET sensors and RootChip technology. New Phytologist 202, 198–208.
Dynamic imaging of cytosolic zinc in Arabidopsis roots combining FRET sensors and RootChip technology.Crossref | GoogleScholarGoogle Scholar | 24372442PubMed |

Leitenmaier B, Küpper H (2013) Compartmentation and complexation of metals in hyperaccumulator plants. Frontiers in Plant Science 4, 374
Compartmentation and complexation of metals in hyperaccumulator plants.Crossref | GoogleScholarGoogle Scholar | 24065978PubMed |

Maret W (2004) Exploring the zinc proteome. Journal of Analytical Atomic Spectrometry 19, 15–19.
Exploring the zinc proteome.Crossref | GoogleScholarGoogle Scholar |

Maret W (2009) Molecular aspects of human cellular zinc homeostasis: redox control of zinc potentials and zinc signals. Biometals 22, 149–157.
Molecular aspects of human cellular zinc homeostasis: redox control of zinc potentials and zinc signals.Crossref | GoogleScholarGoogle Scholar | 19130267PubMed |

Maret W (2012) New perspectives of zinc coordination environments in proteins. Journal of Inorganic Biochemistry 111, 110–116.
New perspectives of zinc coordination environments in proteins.Crossref | GoogleScholarGoogle Scholar | 22196021PubMed |

Maret W (2013) Inhibitory zinc sites in enzymes. Biometals 26, 197–204.
Inhibitory zinc sites in enzymes.Crossref | GoogleScholarGoogle Scholar | 23456096PubMed |

Novikova GV, Mur LA, Nosov AV, Fomenkov AA, Mironov KS, Mamaeva AS, Shilov ES, Rakitin VY, Hall MA (2017) Nitric oxide has a concentration-dependent effect on the cell cycle acting via EIN2 in Arabidopsis thaliana cultured cells. Frontiers in Physiology 8, 142
Nitric oxide has a concentration-dependent effect on the cell cycle acting via EIN2 in Arabidopsis thaliana cultured cells.Crossref | GoogleScholarGoogle Scholar | 28344560PubMed |

Nowakowski AB, Petering DH (2011) Reactions of the fluorescent sensor, Zinquin, with the zinc-proteome: adduct formation and ligand substitution. Inorganic Chemistry 50, 10124–10133.
Reactions of the fluorescent sensor, Zinquin, with the zinc-proteome: adduct formation and ligand substitution.Crossref | GoogleScholarGoogle Scholar | 21905645PubMed |

Nowakowski A, Karim M, Petering D (2015) Zinc proteomics. In ‘Encyclopedia of inorganic and bioinorganic chemistry’. (Ed. R Scott) pp. 1–10. (John Wiley & Sons: Hoboken, NJ, USA)

Oteiza PI (2012) Zinc and the modulation of redox homeostasis. Free Radical Biology & Medicine 53, 1748–1759.
Zinc and the modulation of redox homeostasis.Crossref | GoogleScholarGoogle Scholar |

Peroza EA, dos Santos Cabral A, Wan X, Freisinger E (2013) Metal ion release from metallothioneins: proteolysis as an alternative to oxidation. Metallomics 5, 1204–1214.
Metal ion release from metallothioneins: proteolysis as an alternative to oxidation.Crossref | GoogleScholarGoogle Scholar | 23835914PubMed |

Petering DH, Mahim A (2017) Proteomic high affinity Zn2+ trafficking: where does metallothionein fit in? International Journal of Molecular Sciences 18, 1289
Proteomic high affinity Zn2+ trafficking: where does metallothionein fit in?Crossref | GoogleScholarGoogle Scholar |

Qin Y, Dittmer PJ, Park JG, Jansen KB, Palmer AE (2011) Measuring steady-state and dynamic endoplasmic reticulum and Golgi Zn2+ with genetically encoded sensors. Proceedings of the National Academy of Sciences of the United States of America 108, 7351–7356.
Measuring steady-state and dynamic endoplasmic reticulum and Golgi Zn2+ with genetically encoded sensors.Crossref | GoogleScholarGoogle Scholar | 21502528PubMed |

Rana U, Kothinti R, Meeusen J, Tabatabai NM, Krezoski S, Petering DH (2008) Zinc binding ligands and cellular zinc trafficking: apo-metallothionein, glutathione, TPEN, proteomic zinc, and Zn-Sp1. Journal of Inorganic Biochemistry 102, 489–499.
Zinc binding ligands and cellular zinc trafficking: apo-metallothionein, glutathione, TPEN, proteomic zinc, and Zn-Sp1.Crossref | GoogleScholarGoogle Scholar | 18171589PubMed |

Ricachenevsky FK, Menguer PK, Sperotto RA, Fett JP (2015) Got to hide your Zn away: molecular control of Zn accumulation and biotechnological applications. Plant Science 236, 1–17.
Got to hide your Zn away: molecular control of Zn accumulation and biotechnological applications.Crossref | GoogleScholarGoogle Scholar | 26025516PubMed |

Richard O, Pineau C, Loubet S, Chalies C, Vile D, Marquès L, Berthomieu P (2011) Diversity analysis of the response to Zn within the Arabidopsis thaliana species revealed a low contribution of Zn translocation to Zn tolerance and a new role for Zn in lateral root development. Plant, Cell & Environment 34, 1065–1078.
Diversity analysis of the response to Zn within the Arabidopsis thaliana species revealed a low contribution of Zn translocation to Zn tolerance and a new role for Zn in lateral root development.Crossref | GoogleScholarGoogle Scholar |

Sarret G, Willems G, Isaure MP, Marcus MA, Fakra SC, Frérot H, Pairis S, Geoffroy N, Manceau A, Saumitou‐Laprade P (2009) Zinc distribution and speciation in Arabidopsis halleri × Arabidopsis lyrata progenies presenting various zinc accumulation capacities. New Phytologist 184, 581–595.
Zinc distribution and speciation in Arabidopsis halleri × Arabidopsis lyrata progenies presenting various zinc accumulation capacities.Crossref | GoogleScholarGoogle Scholar | 19761446PubMed |

Schenk RU, Hildebrandt AC (1972) Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Canadian Journal of Botany 50, 199–204.
Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures.Crossref | GoogleScholarGoogle Scholar |

Sinclair SA, Senger T, Talke IN, Cobbett CS, Haydon MJ, Kraemer U (2018) Systemic upregulation of MTP2-and HMA2-mediated Zn partitioning to the shoot supplements local Zn deficiency responses. The Plant Cell 30, 2463–2479.
Systemic upregulation of MTP2-and HMA2-mediated Zn partitioning to the shoot supplements local Zn deficiency responses.Crossref | GoogleScholarGoogle Scholar | 30150315PubMed |

Soshinkova TN, Radyukina NL, Korolkova DV, Nosov AV (2013) Proline and functioning of the antioxidant system in Thellungiella salsuginea plants and cultured cells subjected to oxidative stress. Russian Journal of Plant Physiology 60, 41–54.
Proline and functioning of the antioxidant system in Thellungiella salsuginea plants and cultured cells subjected to oxidative stress.Crossref | GoogleScholarGoogle Scholar |

Srivastava PK, Anand A (2015) The inhibitory effect of metals and other ions on acid phosphatase activity from Vigna aconitifolia seeds. Preparative Biochemistry & Biotechnology 45, 33–41.
The inhibitory effect of metals and other ions on acid phosphatase activity from Vigna aconitifolia seeds.Crossref | GoogleScholarGoogle Scholar |

Vera-Estrella R, Miranda-Vergara MC, Barkla BJ (2009) Zinc tolerance and accumulation in stable cell suspension cultures and in vitro regenerated plants of the emerging model plant Arabidopsis halleri (Brassicaceae). Planta 229, 977–986.
Zinc tolerance and accumulation in stable cell suspension cultures and in vitro regenerated plants of the emerging model plant Arabidopsis halleri (Brassicaceae).Crossref | GoogleScholarGoogle Scholar | 19148674PubMed |

Wilson M, Hogstrand C, Maret W (2012) Picomolar concentrations of free zinc (II) ions regulate receptor protein-tyrosine phosphatase β activity. Journal of Biological Chemistry 287, 9322–9326.
Picomolar concentrations of free zinc (II) ions regulate receptor protein-tyrosine phosphatase β activity.Crossref | GoogleScholarGoogle Scholar | 22275360PubMed |

Yamasaki S, Hasegawa A, Hojyo S, Ohashi W, Fukada T, Nishida K, Hirano T (2012) A novel role of the L-type calcium channel α1D subunit as a gatekeeper for intracellular zinc signaling: zinc wave. PLoS One 7, e39654
A novel role of the L-type calcium channel α1D subunit as a gatekeeper for intracellular zinc signaling: zinc wave.Crossref | GoogleScholarGoogle Scholar | 22745805PubMed |

Zalewski P, Truong-Tran A, Lincoln S, Ward D, Shankar A, Coyle P, Jayaram L, Copley A, Grosser D, Murgia C, Lang C, Ruffin R (2006) Use of a zinc fluorophore to measure labile pools of zinc in body fluids and cell-conditioned media. BioTechniques 40, 509–520.
Use of a zinc fluorophore to measure labile pools of zinc in body fluids and cell-conditioned media.Crossref | GoogleScholarGoogle Scholar | 16629398PubMed |

Zlobin IE, Kartashov AV, Shpakovski GV (2017) Different roles of glutathione in copper and zinc chelation in Brassica napus roots. Plant Physiology and Biochemistry 118, 333–341.
Different roles of glutathione in copper and zinc chelation in Brassica napus roots.Crossref | GoogleScholarGoogle Scholar | 28683402PubMed |