Využitie akustickej metódy na sledovanie interakcií cytochrómu c s aptamérom-modifikovanými nanodrôtmi

Využitie akustickej metódy na sledovanie interakcií cytochrómu c s aptamérom-modifikovanými nanodrôtmi

Sekcia: 
Rok:
2022

Celkové hodnotenie

Vedecká práca
67%
Prevedenie (dizajn)
67%
Diskusná interakcia
PoužívateľVedecká prácaDizajnDiskusná interakcia
Doc. RNDr. Iveta Waczulíková PhD.100%100%-

Využitie akustickej metódy na sledovanie interakcií cytochrómu c s aptamérom-modifikovanými nanodrôtmi

Martin Csiba1 , Marek Tatarko , Tibor Hianik , Veronika Šubjaková ,
1 Katedra jadrovej fyziky a biofyziky FMFI UK
csiba17@uniba.sk

In this work, we focused on the study of the interaction of cytochrome c (cyt c) with gold nanowires (AuNWs) modified with DNA aptamer specific for cyt c (NH2-Apt-cytc). Interactions of Apt-Cyt  or AuNWs modified with Apt-Cyt and cyt c embedded in lipid membranes formed by DMPC (1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine) and DMPG (1,2-Dimyristoyl-sn-glycero-3-phosphorylglycerol) in a molar ratio of 1 : 1 were studied by the acoustic method of Quartz Crystal Microbalance with Dissipation (QCM-D). Lipid layers of DMPC and DMPG were formed on a self-assembled layer of 1-dodecanethiol adsorbed on the gold surface of quartz crystals. We optimized the properties of the layers using buffers with concentrations of salts (MgCl2 and CaCl2). We verified the interaction by the acoustic method QCM-D at a flow rate of 50 ml / min, when we found differences between functionalized and non-functionalized AuNWs. To verify the specificity of the interaction, we also applied DNA aptamer which was non-specific for cyt c. The non-specific aptamer for our measurements was the penicillin-specific DNA aptamer. When using it, we did not detect any interaction with the layers.

Poďakovanie: 

This work was funded by the European Union's Horizon 2020 Research and Innovation Program, program H2020-MSCA-RISE-2020, grant number 101007299 and the VEGA Scientific Grant Agency, project number 1/0419/20.

Zdroje: 

HUELLEMEIER, Holly A., Necla M. EREN a Joana ORTEGA-ANAYA, 2022. Application of quartz crystal microbalance with dissipation (QCM-D) to study low-temperature adsorption and fouling of milk fractions on stainless steel. Chemical Engineering Science [online]. 2022, roč. 247, s. 117004. ISSN 0009-2509. Dostupné na: doi:10.1016/J.CES.2021.117004

KANAZAWA, K. Keiji a Joseph G. GORDON, 1985. Frequency of a Quartz Microbalance in Contact with Liquid. Analytical Chemistry [online]. 1985, roč. 57, č. 8, s. 1770–1771. ISSN 15206882. Dostupné na: doi:10.1021/AC00285A062

KELLER, C. A., K. GLASMÄSTAR, V. P. ZHDANOV a B. KASEMO, 2000. Formation of Supported Membranes from Vesicles. Physical Review Letters [online]. 2000, roč. 84, č. 23, s. 5443–5446. ISSN 00319007. Dostupné na: doi:10.1103/PHYSREVLETT.84.5443

LAUDADIO, Emiliano, Cristina MINNELLI, Adolfo AMICI, Luca MASSACCESI, Giovanna MOBBILI a Roberta GALEAZZI, 2018. Liposomal formulations for an efficient encapsulation of epigallocatechin-3-gallate: An in-silico/experimental approach. Molecules [online]. 2018, roč. 23, č. 2. ISSN 14203049. Dostupné na: doi:10.3390/MOLECULES23020441

MANICKAM, Pandiaraj, Ajeet KAUSHIK, Chandran KARUNAKARAN a Shekhar BHANSALI, 2017. Recent advances in cytochrome c biosensing technologies. Biosensors and Bioelectronics [online]. 2017, roč. 87, s. 654–668. ISSN 0956-5663. Dostupné na: doi:10.1016/J.BIOS.2016.09.013

MILOREY, Bridget, Reinhard SCHWEITZER-STENNER, Raghed KURBAJ a Dmitry MALYSHKA, 2019. PH-Induced Switch between Different Modes of Cytochrome c Binding to Cardiolipin-Containing Liposomes. ACS Omega [online]. 2019, roč. 4, č. 1, s. 1386–1400. ISSN 24701343. Dostupné na: doi:10.1021/ACSOMEGA.8B02574

SCHOEFFLER, Allyn J., Carmen R. RUIZ, Allison M. JOUBERT, Xuemei YANG a Vince J. LICATA, 2003. Salt modulates the stability and lipid binding affinity of the adipocyte lipid-binding protein. Journal of Biological Chemistry [online]. 2003, roč. 278, č. 35, s. 33268–33275. ISSN 00219258. Dostupné na: doi:10.1074/JBC.M304955200

SCHWEITZER-STENNER, Reinhard, 2018. Relating the multi-functionality of cytochrome c to membrane binding and structural conversion. Biophysical Reviews [online]. 2018, roč. 10, č. 4, s. 1151–1185. ISSN 18672469. Dostupné na: doi:10.1007/S12551-018-0409-4

SUBJAKOVA, Veronika, Veronika ORAVCZOVA a Tibor HIANIK, 2021. Polymer Nanoparticles and Nanomotors Modified by DNA/RNA Aptamers and Antibodies in Targeted Therapy of Cancer. polymers [online]. 2021 [cit. 10.3.2021]. Dostupné na: doi:10.3390/polym13030341

WIŚNIEWSKA-BECKER, Anna, 2013. Biomembrane models. V: Drug-Biomembrane Interaction Studies: The Application of Calorimetric Techniques [online]. B.m.: Elsevier Inc., s. 47–95. ISBN 9781907568053. Dostupné na: doi:10.1533/9781908818348.47

YEAGLE, Philip L., 2016. Membrane Proteins. V: The Membranes of Cells [online]. B.m.: Elsevier, s. 219–268 [cit. 8.5.2021]. Dostupné na: doi:10.1016/B978-0-12-800047-2.00010-3

ZHANG, Yabin, Ke YUAN a Li ZHANG, 2019. Micro/Nanomachines: from Functionalization to Sensing and Removal [online]. 1. apríl 2019. B.m.: Wiley-Blackwell. [cit. 30.3.2021]. Dostupné na: doi:10.1002/admt.201800636

ZIMA, J a P ELEDER, 2012. Metoda QCM-D dobývá jeden obor za druhým. Chemagazín [online]. 2012, roč. 22, č. 5 [cit. 11.4.2022]. Dostupné na: https://www.chromspec.cz/download/chromspec-qcm-d.pdf

 

Diskusia

Ďakujeme za pekný príspevok z jednej z najmodernejších oblastí biofyzikálneho výskumu nanosystémov a ich aplikácií. Môžete nám prezradiť, ktorý krok v experimentálnom protokole považujete za najnáročnejší a prečo?

iwa

Ďakujem za zaujímavý prispevok z oblasti nanotechnológií. Chcem sa opýtať, akú úlohu hrá výber tvaru nanočastic. Prečo ste vybrali nanodôty. Majú nejakú výhodu oproti napr. sférickým nanočasticiam? MZ