Metal ion binding to apo, holo, and reconstituted horse spleen ferritin.
Abstract: The binding of Cd2+, Zn2+, Cu2+, Ni2+, Co2+, Mn2+, and Mg2+ to apo, holo, reconstituted horse spleen ferritin (HoSF), and native holo HoSF with phosphate removed was measured by gel-exclusion chromatography. Three classes of strong binding interactions (Kd < 10(-7) M) with apo HoSF at pH 7.5 were found for the various M2+ studied: high stoichiometric binding (30-54 M2+/HoSF) for Cd2+, Zn2+, Cu2+, with two protons released per metal bound; intermediate binding (16 M2+/HoSF) for Ni2+ and Co2+, with one proton released per metal bound; and low levels of binding (2-12 M2+/HoSF) for Mn2+, Mg2+, and Fe2+, with < 0.5 protons released per metal bound. M2+ binding to apo HoSF was nearly abolished at pH 5.5, except for Fe2+ and Cu2+, which remained unaffected by pH alteration. Holo HoSF bound much higher levels of M2+, a result directly attributable to the presence of phosphate binding sites. This conclusion was confirmed by decreased binding of M2+ to HoSF reconstituted in the absence of phosphate and by native holo HoSF with phosphate chemically removed. The binding of Cd2+ to apo HoSF was 54 per HoSF, but in the presence of developing core, the amount bound decreased to about 30 Cd2+/HoSF. This result indicated that Cd2+ and developing core were competing for the same sites on the HoSF interior, suggesting that 24 of the Cd2+ were bound to the inside surface. No other M2+ studied bound to the interior of HoSF by this criterion. Several of the M2+ appeared to bind strongly to the phosphate-free mineral core surface in reconstituted HoSF.
Publication Date: 1995-07-01 PubMed ID: 7782791DOI: 10.1016/0162-0134(94)00050-kGoogle Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This study examines how different metal ions interact with three forms of horse spleen ferritin. It was found that cadmium, zinc, and copper had the highest binding levels, while manganese, magnesium, and iron exhibited lower levels. The binding ability of most metal ions significantly decreased at a lower pH, with the notable exceptions of iron and copper. The amount of phosphate present also notably influenced the binding of the metal ions.
Methodology
- The researchers measured the binding of several metal ions (Cd2+, Zn2+, Cu2+, Ni2+, Co2+, Mn2+, Mg2+) to different forms of horse spleen ferritin (apo, holo, reconstituted) and native holo ferritin with phosphate removed. The binding was measured using gel-exclusion chromatography, a technique that separates molecules based on their size.
Main Findings
- For the various metal ions studied, three types of strong binding interactions were found with the apo Horse Spleen Ferritin (HoSF) at a pH of 7.5. The metal ions released different numbers of protons when bound, affecting pH levels.
- Cadmium, zinc, and copper had high binding levels, releasing two protons per metal bound.
- Nickel and cobalt showed intermediate binding, releasing one proton per metal ion bound.
- Meanwhile, manganese, magnesium, and iron demonstrated low levels of binding, releasing less than 0.5 protons per metal ion bound.
- The binding ability of these metal ions to the apo HoSF significantly decreased when the pH was lowered to 5.5, except for iron and copper, which remained unaffected by the pH alteration.
Influence of Phosphate
- Phosphate presence in HoSF had a considerable effect on metal ion binding. The holo form of HoSF showed a much higher level of metal ion binding ability, a result directly attributed to the presence of phosphate binding sites.
- This was further confirmed when less metal ion binding was witnessed in HoSF reconstituted without the presence of phosphate and when native holo HoSF had phosphate chemically removed.
Binding to Ferritin Interior
- The study found that cadmium alone among the tested metal ions was bound to the inside surface of the HoSF. The binding of cadmium to apo HoSF was 54 per ferritin molecule, but this number decreased when a ferritin ‘core’ was developing. This suggests that cadmium and the ferritin core compete for the same binding sites.
- No other tested metal ions appeared to bind to the interior of the HoSF according to this criterion.
Conclusions
- The researchers concluded that several of the tested metal ions appeared to bind strongly to the phosphate-free mineral core surface in reconstituted HoSF. This finding provides more insights into the role of both pH levels and phosphate presence in the binding ability of different metal ions to horse spleen ferritin.
Cite This Article
APA
Pead S, Durrant E, Webb B, Larsen C, Heaton D, Johnson J, Watt GD.
(1995).
Metal ion binding to apo, holo, and reconstituted horse spleen ferritin.
J Inorg Biochem, 59(1), 15-27.
https://doi.org/10.1016/0162-0134(94)00050-k Publication
Researcher Affiliations
- Department of Chemistry Undergraduate Research Program, Brigham Young University, Provo, UT 84602, USA.
MeSH Terms
- Animals
- Apoferritins / metabolism
- Binding Sites
- Binding, Competitive
- Cations, Divalent / metabolism
- Chromatography, Gel
- Ferritins / metabolism
- Horses
- Iron / metabolism
- Oxidation-Reduction
- Phosphates / pharmacology
- Protons
- Spleen / chemistry
Grant Funding
- 5RO1 DK36799-05 / NIDDK NIH HHS
Citations
This article has been cited 14 times.- Han Z, Guo AX, Luo T, Cai T, Mirkin CA. Biomineralization of semiconductor quantum dots using DNA-functionalized protein nanoreactors. Sci Adv 2025 May 9;11(19):eadv6906.
- Cosottini L, Giachetti A, Guerri A, Martinez-Castillo A, Geri A, Zineddu S, Abrescia NGA, Messori L, Turano P, Rosato A. Structural Insight Into a Human H Ferritin@Gold-Monocarbene Adduct: Aurophilicity Revealed in a Biological Context. Angew Chem Int Ed Engl 2025 Jul 21;64(30):e202503778.
- Tricase A, Alhenaki B, Marchianò V, Torsi L, Gupta R, Bollella P. Bioelectrochemically triggered apoferritin-based bionanoreactors: synthesis of CdSe nanoparticles and monitoring with leaky waveguides. Nanoscale Adv 2024 Jan 16;6(2):516-523.
- Huo C, Ming T, Wu Y, Huan H, Qiu X, Lu C, Li Y, Zhang Z, Han J, Su X. Structural and Biochemical Characterization of Silver/Copper Binding by Dendrorhynchus zhejiangensis Ferritin. Polymers (Basel) 2023 Mar 3;15(5).
- Slobodian MR, Petahtegoose JD, Wallis AL, Levesque DC, Merritt TJS. The Effects of Essential and Non-Essential Metal Toxicity in the Drosophila melanogaster Insect Model: A Review. Toxics 2021 Oct 16;9(10).
- Imran Din M, Rafique F, Hussain MS, Arslan Mehmood H, Waseem S. Recent developments in the synthesis and stability of metal ferrite nanoparticles. Sci Prog 2019 Mar;102(1):61-72.
- Ding H, Zhang D, Chu S, Zhou J, Su X. Screening and structural and functional investigation of a novel ferritin from Phascolosoma esculenta. Protein Sci 2017 Oct;26(10):2039-2050.
- Volatron J, Kolosnjaj-Tabi J, Javed Y, Vuong QL, Gossuin Y, Neveu S, Luciani N, Hémadi M, Carn F, Alloyeau D, Gazeau F. Physiological Remediation of Cobalt Ferrite Nanoparticles by Ferritin. Sci Rep 2017 Jan 9;7:40075.
- Wang Z, Huang P, Jacobson O, Wang Z, Liu Y, Lin L, Lin J, Lu N, Zhang H, Tian R, Niu G, Liu G, Chen X. Biomineralization-Inspired Synthesis of Copper Sulfide-Ferritin Nanocages as Cancer Theranostics. ACS Nano 2016 Mar 22;10(3):3453-60.
- He D, Marles-Wright J. Ferritin family proteins and their use in bionanotechnology. N Biotechnol 2015 Dec 25;32(6):651-7.
- Li C, Li Z, Li Y, Zhou J, Zhang C, Su X, Li T. A ferritin from Dendrorhynchus zhejiangensis with heavy metals detoxification activity. PLoS One 2012;7(12):e51428.
- Michon J, Frelon S, Garnier C, Coppin F. Determinations of uranium(VI) binding properties with some metalloproteins (transferrin, albumin, metallothionein and ferritin) by fluorescence quenching. J Fluoresc 2010 Mar;20(2):581-90.
- Huang HQ, Lin QM, Lou ZB. Construction of a ferritin reactor: an efficient means for trapping various heavy metal ions in flowing seawater. J Protein Chem 2000 Aug;19(6):441-7.
- Levi S, Santambrogio P, Corsi B, Cozzi A, Arosio P. Evidence that residues exposed on the three-fold channels have active roles in the mechanism of ferritin iron incorporation. Biochem J 1996 Jul 15;317 ( Pt 2)(Pt 2):467-73.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
