Uptake of Cell-Penetrating Peptide RL2 by Human Lung Cancer Cells: Monitoring by Electron Paramagnetic Resonance and Confocal Laser Scanning Microscopy

Uptake of Cell-Penetrating Peptide RL2 by Human Lung Cancer Cells: Monitoring by Electron Paramagnetic Resonance and Confocal Laser Scanning Microscopy Full article

Journal

Molecules
, E-ISSN: 1420-3049

Output data

Year: 2021, Volume: 26, Number: 18, Article number : 5442, Pages count : DOI: 10.3390/molecules26185442

Tags

cell penetrating peptide; electron spin resonance spectroscopy; confocal microscopy

Authors

Ovcherenko Sergey S. ¹ , Chinak Olga A. ² , Chechushkov Anton ² , Dobrynin Sergey A. ¹ , Kirilyuk Igor A. ¹ , Krumkacheva Olesya A. ³ , Richter Vladimir A. ² , Bagryanskaya Elena G. ¹

Affiliations

1	(Данные Web of science) NN Vorozhtsov Novosibirsk Inst Organ Chem SB RAS, Novosibirsk 630090, Russia
2	(Данные Web of science) Inst Chem Biol & Fundamental Med SB RAS, Novosibirsk 630090, Russia
3	(Данные Web of science) Int Tomog Ctr SB RAS, Novosibirsk 630090, Russia

RL2 is a recombinant analogue of a human kappa-casein fragment, capable of penetrating cells and inducing apoptosis of cancer cells with no toxicity to normal cells. The exact mechanism of RL2 penetration into cells remains unknown. In this study, we investigated the mechanism of RL2 penetration into human lung cancer A549 cells by a combination of electron paramagnetic resonance (EPR) spectroscopy and confocal laser scanning microscopy. EPR spectra of A549 cells incubated with RL2 (sRL2) spin-labeled by a highly stable 3-carboxy-2,2,5,5-tetraethylpyrrolidine-1-oxyl radical were found to contain three components, with their contributions changing with time. The combined EPR and confocal-microscopy data allowed us to assign these three forms of sRL2 to the spin-labeled protein sticking to the membrane of the cell and endosomes, to the spin-labeled protein in the cell interior, and to spin labeled short peptides formed in the cell because of protein digestion. EPR spectroscopy enabled us to follow the kinetics of transformations between different forms of the spin-labeled protein at a minimal spin concentration (3-16 mu M) in the cell. The prospects of applications of spin-labeled cell-penetrating peptides to EPR imaging, DNP, and magnetic resonance imaging are discussed, as is possible research on an intrinsically disordered protein in the cell by pulsed dipolar EPR spectroscopy.

Ovcherenko S.S. , Chinak O.A. , Chechushkov A. , Dobrynin S.A. , Kirilyuk I.A. , Krumkacheva O.A. , Richter V.A. , Bagryanskaya E.G.
Uptake of Cell-Penetrating Peptide RL2 by Human Lung Cancer Cells: Monitoring by Electron Paramagnetic Resonance and Confocal Laser Scanning Microscopy
Molecules. 2021. V.26. N18. 5442 . DOI: 10.3390/molecules26185442 WOS Scopus РИНЦ OpenAlex

Web of science:	WOS:000701981500001
Scopus:	2-s2.0-85114649548
Elibrary:	47026525
OpenAlex:	W3197803668

DB	Citing
Web of science	17
Scopus	17
Elibrary	19
OpenAlex	18