Abstract
The bioactivity of the natural ultrafine carbon form shungite nanocarbon (ShC) is of particular interest both for biomedical applications of such nanomaterials and their negative impact on the aquatic environmental. Here we studied the interaction of serum albumin (SA) with ShC nanoparticles in aqueous dispersion with respect to its structural-dynamic, thermodynamic, and hydrodynamic effects. Electron spin resonance (EPR) with a 5-DOXYL-stearic acid spin probe (5DSA) demonstrates that ShC can affect fatty acid (FA) binding by SA, protein conformation in the stearic FA spin probe binding region, and protein aggregation due to the partial transfer of FA to the ShC nanoparticles. The ratio of SA fractions changes in the presence of ShC in favor of the fraction that is less saturated with FA as shown by differential scanning calorimetry (DSC). The stability of interaction with ShC is significantly higher for aggregates of SA molecules that carry physiological amounts of FA, compared to aggregates of the FA-free protein, as studied by dynamic light scattering (DLS) analysis. Generally, the mixed dispersion of SA and ShC nanoparticles is more homogeneous than the SA solution alone. This is manifested both in the size of the molecular associates and in the microenvironment of the protein-bound FA. The formation of the SA–ShC interface is likely to result in a greater uniformity of the FA binding sites and a decrease in protein fractions and "hot patches" on the protein surface responsible for the supramolecular heterogeneity of the protein in solution.
Similar content being viewed by others
References
Arcioni A, Bacchiocchi C, Grossi L, Nicolini A, Zannoni C (2002) Electron spin resonance studies of order and dynamics in a nematic liquid crystal containing a dispersed aerosol. J Phys Chem B 106:9245–9251. https://doi.org/10.1021/jp020554j
Augustyniak-Jablokow MA, Yablokov YV, Andrzejewski B, Kempiński W, Łoś S, Tadyszak K, Yablokov MY, Zhikharev VA (2010) EPR and magnetism of the nanostructured natural carbonaceous material shungite. Phys Chem Minerals 37:237–247. https://doi.org/10.1007/s00269-009-0328-9
Brewer SH, Glomm WR, Johnson MC, Knaq MK, Franzen S (2005) Probing BSA binding to citrate-coated gold nano-particles and surfaces. Langmuir 21:9303–9307. https://doi.org/10.1021/la050588t
Budyka MF, Sheka EF, Popova NA (2017) Graphene quantum dots: theory and experiment. Rev Adv Mater Sci 51:35–49
Chakraborty S, Joshi P, Shanker V, Ansari ZA, Singh SP, Chakrabarti P (2011) Contrasting effect of gold nanoparticles and nanorods with different surface modifications on the structure and activity of bovine serum albumin. Langmuir 27(12):7722–7731. https://doi.org/10.1021/la200787t
Chen Z, Wu D (2012) Spectroscopic studies on the interaction between ZnSe nanoparticles with bovine serum albumin. J Lumin 132:2968–2974. https://doi.org/10.1016/j.jlumin.2012.06.028
Chou NH, Pierce N, Lei Y, Perea-López N, Fujisawa R, Subramanian S, Robinson JA, Chen G, Omichi K, Rozhkov SS, Rozhkova NN, Terrones M, Harutyunyan AR (2018) Carbon-rich shungite as a natural resource for efficient Li-ion battery electrodes. Carbon 130:105–111. https://doi.org/10.1016/j.carbon.2017.12.109
Chumakova NA, Rebrikova AT, Talyzin AV, Korobov MV (2018) Properties of graphite oxide powders and membranes as revealed by electron paramagnetic resonance spectroscopy. J Phys Chem C 122:22750–22759. https://doi.org/10.1021/acs.jpcc.8b07221
Couto C, Vitorino R, Daniel-da-Silva AL (2017) Gold nanoparticles and bioconjugation: a pathway for proteomic applications. Crit Rev Biotechnol 37:238–250. https://doi.org/10.3109/07388551.2016.1141392
Dhami AK, Bhat S, Sharma A, Bhat SV (2008) Spin probe ESR studies of dynamics of single walled carbon nanotubes. Spectrochim Acta (Part A) 69:1178–1182. https://doi.org/10.1016/j.saa.2007.06.034
Esfandfar P, Falahati M, Saboury A (2016) Spectroscopic studies of interaction between CuO nanoparticles and bovine serum albumin. J Biomol Struct Dyn 34:1962–1968. https://doi.org/10.1080/07391102.2015.1096213
Goryunov AS, Borisova AG (2014) Probable mechanism of haemoglobin autoxidation in water dispersions of carbon based nanomaterials. Trans Karelian Res Centre RAS 5:71–75 (in Russian)
Gossmann R, Fahrländer E, Hummel M, Mulaca D, Brockmeyerb J, Langer K (2015) Comparative examination of adsorption of serum proteins on HSA- and PLGA-based nanoparticles using SDS-PAGE and LC-MS. Eur J Pharm Biopharm 93:80–87. https://doi.org/10.1016/j.ejpb.2015.03.021
Gurachevsky A, Shimanovitch E, Gurachevskaya T, Muravsky V (2007) Intra-albumin migration of bound fatty acid probed by spin label ESR. Biochem Biophys Res Commun 360:852–856. https://doi.org/10.1016/j.bbrc.2007.06.140
Hairulin AR, Stepanova TP, Rozhkova NN, Gladchenko SN (2012a) Static dielectric polarization of shungite carbon structural elements in benzene series solvents. Phys Math 4(153):111–114 (in Russian)
Hairulin AR, Stepanova TP, Rozhkova NN, Gladchenko SN (2012b) Dipole moments of fullerene C60 in benzene, toluene and orthoxylene. Phys Math 3(153):81–89 (in Russian)
Huang R, Carney RP, Ikuma K, Stellacci F, Lau BL (2014) Effects of surface compositional and structural heterogeneity on nanoparticle–protein interactions: different protein configurations. ACS Nano 8:5402–5412. https://doi.org/10.1021/nn501203k
Junk MJN, Spiess HW, Hinderberger D (2011) DEER in biological multispin-systems: a case study on the fatty acid binding to human serum albumin. J Magn Reson 210:210–217. https://doi.org/10.1016/j.jmr.2011.03.003
Kempinski M, Łos S, Florczak P, Kempinski W, Jurga S (2017) EPR and impedance measurements of graphene oxide and reduced graphene oxide. Acta Physica Polonica A 132:81–85. https://doi.org/10.12693/APhysPolA.132.81
Likhtenstein GI (1976) Spin-labeling methods in molecular biology. Wiley, London
Lin HA, Sato Y, Segawa Y, Nishihara T, Sugimoto N, Scott LT, Higashiyama T, Itami KA (2018) Water-soluble warped nanographene: synthesis and applications for photoinduced cell death. Angew Chem Int Ed Engl 57:2874–2878. https://doi.org/10.1002/anie.201713387
Liu J, Fu S, Yuan B, Li Y, Deng Z (2010) Toward a universal “adhesive nanosheet” for the assembly of multiple nanoparticles based on a protein-induced reduction/decoration of graphene oxide. J Am Chem Soc 132:7279–7281. https://doi.org/10.1021/ja100938r
Mandal S, Hossain M, Devi PS, Kumar GS, Chaudhuri K (2013) Interaction of carbon nanoparticles to serum albumin: elucidation of the extent of perturbation of serum albumin conformations and thermodynamical parameters. J Hazard Mater 248–249:238–245. https://doi.org/10.1016/j.jhazmat.2013.01.009
Maruyama T, Katoh S, Nakajima M, Nabetani H (2001) Mechanism of bovine serum albumin aggregation during ultrafiltration. Biotechnol Bioeng 75:233–238. https://doi.org/10.1002/bit.10001
Militello V, Vetri V, Leone M (2003) Conformational changes involved in thermal aggregation processes of bovine serum albumin. Biophys Chem 105:133–141. https://doi.org/10.1016/S0301-4622(03)00153-4
Morrisett JD, Pownall HJ, Gotto AM, Jr (1975) Bovine serum albumin. Study of the fatty acid and steroid binding sites using spin-labeled lipids. J Biol Chem 250:2487–2494
Muravsky V, Gurachevskaya T, Berezenko S, Schnurra K, Gurachevsky A (2009) Fatty acid binding sites of human and bovine albumins: differences observed by spin probe ESR. Spectrochim Acta Part A Mol Biomol Spectrosc 74:42–47. https://doi.org/10.1016/j.saa.2009.05.003
Ni Y, Zhanga F, Kokot S (2013) Graphene oxide as a nanocarrier for loading and delivery of medicinal drugs and as a biosensor for detection of serum albumin. Anal Chim Acta 769:40–48. https://doi.org/10.1016/j.aca.2013.01.038
Pavićević AA, Popović-Bijelić AD, Mojović MD, Šušnjar SV, Bačić GG (2014) Binding of doxyl stearic spin labels to human serum albumin: an EPR study. J Phys Chem B 118:10898–10905. https://doi.org/10.1021/jp5068928
Peters T (1996) All about albumin Biochemistry, genetics and medical applications. Academic Press, San Diego. https://doi.org/10.1002/food.19970410631
Rajeshwari A, Pakrashi S, Madhumita SD, Iswarya V, Chandrasekaran N, Mukherjee A (2014) Spectroscopic studies on the interaction of bovine serum albumin with Al2O3 nanoparticles. J Lumin 145:859–865. https://doi.org/10.1016/j.jlumin.2013.08.073
Razbirin BS, Rozhkova NN, Sheka EF, Nelson DK, Starukhin AN, Goryunov AS (2014) Spectral properties of shungite quantum dots. Nanosyst Phys Chem Math 5:217–233
Razbirin BS, Rozhkova NN, Sheka EF (2016) Photonics of shungite quantum dots. In: Aliofkhazraei M, et al. (eds) Graphene science handbook: electrical and optical properties, vol 1. CRC Press, Boca Raton, pp 425–436
Reichenwallner J, Oehmichen M-T, Schmelzer CEH, Hauenschild T, Kerth A, Hinderberger D (2018) Exploring the pH-induced functional phase space of human serum albumin by EPR spectroscopy. Magnetochemistry 4:47. https://doi.org/10.3390/magnetochemistry4040047
Roy S, Das TK (2014) Spectroscopic studies of interaction between biologically synthesized silver nanoparticles and bovine serum albumin. J Nanosci Nanotechnol 14:4899–4905. https://doi.org/10.1166/jnn.2014.9508
Rozhkov SP, Goryunov AS (2000) Effects of inorganic salts on the structural heterogeneity of serum albumin solutions. Eur Biophys J 28:639–647
Rozhkov SP, Goryunov AS (2013) Interaction of shungite carbon nanoparticles with blood protein and cell components. Russ J Gen Chem 83:2585–2595. https://doi.org/10.1134/S1070363213130021
Rozhkov SP, Goryunov AS, Rozhkova NN (2007) EPR spin-probe study of carbon nanoparticles hydration properties in aqueous dispersions. In: Vezirogly TE, et al. (eds) Hydrogen materials science and chemistry of carbon nanomaterials. Springer Science+Bisuness Media, Luxembourg, pp 539–544
Rozhkov SP, Goryunov AS, Rozhkov SS (2018) Water dispersions of natural graphene based carbon nanoparticles: ESR spin probe study. Colloids Surf A 537:549–556. https://doi.org/10.1016/j.colsurfa.2017.10.062
Rozhkova NN (2013) Aggregation and stabilization of shungite carbon nanoparticles. Russ J Gen Chem 83:2676–2685. https://doi.org/10.1134/S1070363213130136
Rozhkova NN, Rozhkov SP, Goryunov AS (2016) Natural graphene based shungite nanocarbon. In: Sattler KD (eds) Carbon nanomaterials sourcebook. Graphene, fullerenes, nanotubes, and nanodiamonds, vol 1. CRC Press, Boca Raton, pp 153–176. https://doi.org/10.1201/b19679
Sajo MEJ, Kim CS, Kim SK, Shim KY, Kang TY, Lee KJ (2017) Antioxidant and anti-inflammatory effects of shungite against ultraviolet B irradiation-induced skin damage in hairless mice. Oxid Med Cell Longev 2017:7340143. https://doi.org/10.1155/2017/7340143
Sancataldo G, Vetri V, Foderà V, Di Cara G, Militello V, Leone M (2014) Oxidation enhances human serum albumin thermal stability and changes the routes of amyloid fibril formation. PLoS One 9:e84552. https://doi.org/10.1371/journal.pone.0084552
Sheka EF, Rozhkova NN (2014) Shungite as the natural pantry of nanoscale reduced graphene oxide. Int J Smart Nano Mater 5:1–16. https://doi.org/10.1080/19475411.2014.885913
Sheka EF, Rozhkova NN, Hołderna-Natkaniec K, Natkaniec I (2014) Neutron scattering study of reduced graphene oxide of natural origin. JETP Lett 99:650–655. https://doi.org/10.1134/S0021364014110113
Sheka EF, Hołderna-Natkaniec K, Natkaniec I, Krawczyk J, Golubev EA, Rozhkova NN, Kim VV, Popova NA, Popova VA (2019) Computationally supported neutron scattering study of natural and synthetic amorphous carbons. J Phys Chem C 123:15841–15850. https://doi.org/10.1021/acs.jpcc.9b03675
Sun H, Zhao A, Gao N, Li K, Ren J, Qu X (2015) Deciphering a nanocarbon-based artificial peroxidase: chemical identification of the catalytically active and substrate-binding sites on graphene quantum dots. Angew Chem Int Ed 54(24):7176–7180. https://doi.org/10.1002/anie.201500626
Sun B, Zhang Y, Chen W, Wang K, Zhu L (2018) Concentration dependent effects of bovine serum albumin on graphene oxide colloidal stability in aquatic environment. Environ Sci Technol 52:7212–7219. https://doi.org/10.1021/acs.est.7b06218
Sychev MM, Myakin SV, Ogurytsov KA, Rozhkova NN, Vasina ES, Matveichikova PV, Belyaev VV (2017) Luminescence of ZnS: Cu particles modified by shungite nanocarbon. J Opt Technol 84:49–52. https://doi.org/10.1364/JOT.84.000049
Tamburri E, Carcione R, Politi S, Angjellari M, Lazzarini L, Vanzetti LE, Macis S, Pepponi G, Terranova ML (2018) Shungite carbon as unexpected natural source of few-layer graphene platelets in a low oxidation state. Inorg Chem 57:8487–8498. https://doi.org/10.1021/acs.inorgchem.8b01164
Treuel L, Brandholt S, Maffre P, Wiegele S, Shang L, Nienhaus GU (2014) Impact of protein modification on the protein corona on nanoparticles and nanoparticle–cell interactions. ACS Nano 8:503–513. https://doi.org/10.1021/nn405019v
Tsai DH, Del Rio FW, Keene AM, Tyner KM, MacCuspie RI, Cho TJ, Zachariah MR, Hackley VA (2011) Adsorption and conformation of serum albumin protein on gold nanoparticles investigated using dimensional measurements and in situ spectroscopic methods. Langmuir 27:2464–2477. https://doi.org/10.1021/la104124d
Wang X, Guo W, Hu Y, Wu J, Wei H (2016) Nanozymes: next wave of artificial enzymes. Springer, Berlin. https://doi.org/10.1007/978-3-662-53068-9
Wang B, Fielding AJ, Dryfe RAW (2019) Electron paramagnetic resonance investigation of the structure of graphene oxide: pH-dependence of the spectroscopic response. ACS Appl Nano Mater 2:19–27. https://doi.org/10.1021/acsanm.8b01329
Xu Y, Watermann T, Limbach H-H, Gutmann T, Sebastian D, Buntkowsky G (2014) Water and small organic molecules as probes for geometric confinement in well-ordered mesoporous carbon materials. Phys Chem Chem Phys 16:9327–9336. https://doi.org/10.1039/c4cp00808a
Xu ZQ, Yang QQ, Lan JY, Zhang JQ, Peng W, Jin JC, Jiang FL, Liu Y (2016) Interactions between carbon nanodots with human serum albumin and γ-globulins: the effects on the transportation function. J Hazard Mater 301:242–249. https://doi.org/10.1016/j.jhazmat.2015.08.062
Zaman M, Ahmad E, Qadeer A, Rabbani G, Khan RH (2014) Nanoparticles in relation to peptide and protein aggregation. Int J Nanomed 9:899–912. https://doi.org/10.2147/IJN.S54171
Zhao J, Wang Z, White JC, Xing B (2014) Graphene in the aquatic environment: adsorption, dispersion, toxicity and transformation. Environ Sci Technol 48:9995–10009. https://doi.org/10.1021/es5022679
Acknowledgements
The study was carried out under state order (project no. 0221-2017-0044).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Goryunov, A., Rozhkov, S. & Rozhkova, N. Fatty acid transfer between serum albumins and shungite carbon nanoparticles and its effect on protein aggregation and association. Eur Biophys J 49, 85–94 (2020). https://doi.org/10.1007/s00249-019-01414-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00249-019-01414-y