Public Release: 23-Apr-2018
Lomonosov Moscow State University
This is the complex of the Nav1.4 channel from human muscle cells with the Hm-3 toxin extracted from the venom of the Heriaeus melloteei spider. (A) The interaction of Hm-3 (blue/purple) with the first voltage-sensing domain (D1) of the channel (sandy/red) revealed by NMR data. Lateral view from the lipid bilayer. (B) The toxin-channel complex. View of the membrane plane from the extracellular space.
Credit: Alexander Vasilevskiy
A team of Russian scientists together with foreign colleagues found out that the venom of crab spider Heriaeus melloteei may be used as a basis for developing treatment against hypokalemic periodic paralysis. This disease is caused by genetic mutation that leads to the occurrence of the so-called ω-currents or leakage currents via voltage-gated ion channels Nav1.4 in skeletal muscles. As a result of such “leakages” the muscles become unable to respond to the signals sent by the nervous system and a patient suffers muscle weakness up to total paralysis. There is still no reliable medicinal drug to cure all cases of this disease. The results of the work were published in Proceedings of the National Academy of Sciences (PNAS) journal.
Each cell membrane has ion channels – protein pores that selectively conduct ions in and out of the cell. Due to their work membrane became electrically charged and has some transmembrane potential. At rest the potential kept on the same level. But under the influence of various signals certain ion channels may open or close, changing the flow of ions in and out of the cell and the charge of the membrane. As a result, certain cells (neurons, muscle cells, and gland cells) are activated and are able to respond to a signal.
However, sometimes channel-coding genes get damaged and cause inadequate response. For example, defects in certain domains of voltage-gated sodium channels Nav1.4 in muscle cells cause the membrane to “leak out” even when the channels are closed. Sodium ions penetrate the membrane and change the electric potential. In this case signals of the nervous system are not able to activate the muscle cells, and a patient develops paralysis. People with type 2 hypokalemic periodic paralysis suffer from muscle weakness up to total immobilization. Unfortunately, existing medicinal drugs against this condition are often inefficient.
“In our work we studied human voltage-gated ion channels, in particular the mutated forms of Nav1.4 channel from skeletal muscle. These mutations lead to a severe disease, type 2 hypokalemic periodic paralysis. We are the first to prove that there are natural chemical compounds able to block the leakage currents through mutated channels,” explained Mikhail Petrovich Kirpichnikov, Doctor of Biological Sciences, member of the Russian Academy of Sciences, and dean of the Faculty of Biology, MSU.
Using a range of genetic methods, protein engineering, electrophysiology, NMR spectroscopy, and computer modeling, the scientists studied the reasons for abnormal activities in the channels damaged by mutation. For the first time they suggested a blocking agent for leakage currents – a toxin Hm-3 extracted from the venom of Heriaeus melloteei spider. According to the data obtained using site-directed mutagenesis, electrophysiology, NMR spectroscopy, and computer modeling, the toxin fixes the voltage-sensing domain of the channel in the position that prevents leakage of ions.
“The discovery of this toxin property gives us hope of developing efficient medicinal drugs for the treatment of patients with hypokalemic paralysis and other similar diseases. The model of interaction between the channel and the toxin gives us prospects for the development of new drugs,” concludes Alexander Vasilevskiy, Candidate of Chemical Sciences, a lecturer at MSU, and the head of a laboratory at the Institute of Bioorganic Chemistry of the Russian Academy of Sciences.
The structural part of the work was commented on by Zakhar Shenkarev, Doctor of Physical and Mathematical Sciences, Professor of the Russian Academy of Sciences, a lecturer at Moscow Institute of Physics and Technology, and the group leader at the Institute of Bioorganic Chemistry of the Russian Academy of Sciences: “Studying the structure of ion channel complexes in the membrane is a difficult task. The main issue of our work was the instability of samples in the course of the NMR experiment. To solve it we had to come up with a range of new experimental methods that speeded up the process of structural data collection from several days to hours.”
The work was conducted by scientists from Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences together with their colleagues from the Faculty of Biology, MSU, as well as foreign specialists from UCL Institute of Neurology and Johns Hopkins University School of Medicine in Baltimore with the support of the Russian Academy of Sciences (Molecular and Cell Biology Program) and Russian Science Foundation.