Kinetics of DNA condensation with DPPC: effect of calcium and sodium cations
Kinetics of DNA condensation with zwitterionic lipid, dipalmitoylphosphatidylcholine, in the presence of calcium and sodium cations was investigated using fluorescence stopped-flow technique. The process was followed from two points of view: the changes on DNA were monitored with ethidium bromide, and fluorescently labelled phospholipid was used to follow the changes in the lipid bilayer. The measurements showed different reaction kinetics. In the case of the DNA side, a consecutive reaction mechanism was found, while two-step first-order reaction mechanism described well the processes on the lipid side. Both, sodium and calcium cations influence significantly the reaction mechanism. Calcium is necessary to condensate DNA with lipid, while sodium modulates the kinetics of the reactions considerably.
KeywordsLipoplex Reaction kinetics Fluorescent probe Fluorescent label
Authors gratefully acknowledge financial support from the Grant Agency of the Ministry of Education, Science, Research and Sport of the Slovak Republic Grant VEGA 1/0916/16 and from Joint Institute for Nuclear Research in Dubna, Russia JINR Project 04-4-1121-2015/2017.
- Khusainova RS, Dawson KA, Rochev I, Gorelov AV, Ivanitskii GR (1999) Structural changes in DNA-Ca2+-dipalmitoylphosphatidylcholine complexes during changes in the molar ratio of nucleotide/lipid. Microcalorimetric study. Dokl Akad Nauk 367:553–556Google Scholar
- Kovalenko DV, Shafei RA, Zelenina IA, Semenova ML, Samuilova OV, Zhdanov RI (1996) Metallonucleoliposome complexes as vehicle for gene delivery to mouse skeletal muscles in vivo. Genetika 32:1299–1301Google Scholar
- Uhríková D (2014) Divalent metal cations in DNA-phospholipid binding. In: Iglic A, Kulkarni CV (eds) Advances in planar lipid bilayers and liposomes, vol 20. Elsevier, New York, pp 111–136Google Scholar
- Uhríková D, Rapp G, Kunst BH, Balgavý P (1998) Interaction of DNA with DPPC bilayers in the presence of Mg(II) ions. Annual Report of the European Molecular Biology Laboratory, EMBL Outstation at DESY, HamburgGoogle Scholar
- Vojčíková L, Švajdlenka E, Balgavý P (1989) Spin label and microcalorimetric studies of the interaction of DNA with unilamellar phosphatidylcholine liposomes. Gen Physiol Biophys 8:399–406Google Scholar
- Zhdanov RI, Podobed OV, Buneeva OA, Kutsenco NG, Tsvetkova TA, Lavrenova TP (1997) Gene transfer into eukaryotic cells using non-cationic (neutral) liposomes. Vop Med Khim 43:212–213Google Scholar