Recombinant Analogs of Sea Anemone Kunitz-Type Peptides Influence P2X7 Receptor Activity in Neuro-2a Cells

• Published in: Marine drugs Vol. 21; no. 3; p. 192
• Main Authors: Pislyagin, Evgeny A, Menchinskaya, Ekaterina S, Gladkikh, Irina N, Kvetkina, Aleksandra N, Sintsova, Oksana V, Popkova, Darya V, Kozlovskiy, Sergei A, Gorpenchenko, Tatiana Y, Likhatskaya, Galina N, Kaluzhskiy, Leonid A, Ivanov, Alexis S, Andreev, Yaroslav A, Kozlov, Sergey A, Dmitrenok, Pavel S, Aminin, Dmitry L, Leychenko, Elena V
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.03.2023; MDPI
• Subjects: Adenosine Triphosphate - metabolism; Analogs; Animals; Apoptosis; ATP; Binding sites; Ca2+ influx; Calcium; Calcium influx; Cells; Chemical properties; Degeneration; dye uptake; Dyes; Fluorescent dyes; Fluorescent indicators; Health aspects; Immunofluorescence; Kunitz-type peptides; Marine invertebrates; Marine toxins; Membrane proteins; Molecular docking; Molecular Docking Simulation; Molecular weight; Nervous system; Neuroblastoma; Neurodegeneration; Neurons; neuroprotective activity; P2X7 receptor; Peptides; Peptides - chemistry; Physiological aspects; Physiology; Receptors; Receptors, Purinergic P2X7; Recombinant proteins; Recombinants; Sea Anemones - metabolism; Sea-anemones; Signaling; Surface plasmon resonance; Therapeutic targets; Toxicity; Russia; molecular docking; sea anemones; Ca2+ influx; Kunitz-type peptides; dye uptake; P2X7 receptor; neuroprotective activity
• ISSN: 1660-3397; 1660-3397
• DOI: 10.3390/md21030192

Purinergic P2X7 receptors (P2X7) have now been proven to play an important role and represent an important therapeutic target in many pathological conditions including neurodegeneration. Here, we investigated the impact of peptides on purinergic signaling in Neuro-2a cells through the P2X7 subtype in in vitro models. We have found that a number of recombinant peptides, analogs of sea anemone Kunitz-type peptides, are able to influence the action of high concentrations of ATP and thereby reduce the toxic effects of ATP. The influx of calcium, as well as the fluorescent dye YO-PRO-1, was significantly suppressed by the studied peptides. Immunofluorescence experiments confirmed that the peptides reduce the P2X7 expression level in neuronal Neuro-2a cells. Two selected active peptides, HCRG1 and HCGS1.10, were found to specifically interact with the extracellular domain of P2X7 and formed stable complexes with the receptor in surface plasmon resonance experiments. The molecular docking approach allowed us to establish the putative binding sites of the most active HCRG1 peptide on the extracellular domain of the P2X7 homotrimer and propose a mechanism for regulating its function. Thus, our work demonstrates the ability of the Kunitz-type peptides to prevent neuronal death by affecting signaling through the P2X7 receptor.