M2 ion channel

It may be possible to increase the utility of our resources to treat influenza virus infection through combinations of antiviral agents with different modes of action (discussed in Cinatl et al. 2007a De Clercq and Neyts 2007). The sialidase inhibitors, for example, may be able to be used in conjunction with the adamantane-based M2 ion channel inhibitors (Govorkova et al. 2004 Ilyushina et al. 2006), with Ribavirin (Smee et al. 2002) or with non-influenza virus specific therapeutics such as anti-inflammatory drugs (Carter 2007). Combination therapy may also reduce the potential of resistance development (Ilyushina et al. 2006). 

Hagiwara T, Kijima-Suda I, Ido T, Ohrui H, Tomita K (1994) Inhibition of bacterial and viral sialidases by 3-fluoro-V-acetyIneuraminic acid, Carbohydr Res 263 167-172 Haskell TH, Peterson FE, Watson D, Plessas NR, Culbertson T (1970) Neuraminidase inhibition and viral chemotherapy, J Med Chem 13 697-704 Hatakeyama S, Sugaya N, Ito M, Yamazaki M, Ichikawa M, Kimura K, Kiso M, Shimizu H, Kawakami C, Koike K, Mitamura K, Kawaoka Y (2007) Emergence of influenza B viruses with reduced sensitivity to neuraminidase inhibitors, JAMA 297 1435-1442 Hay AJ (1992) The action of adamantanamines against influenza A viruses inhibition of the M2 ion channel protein, Semin Virol 3 21-30... 

A. J. Hay, The action of adamantanamines against influenza A viruses Inhibition of the M2 ion channel protein, Semin. Virol., 3 (1992) 21-30. 

The first step in viral replication and infection (Fig. 17.2) [31] is the interaction between the viral surface HA and sialic acid-containing glycoconjugates on the surface of respiratory tract cells. After binding, the virus is endocytosed, and the low pH of the endosome triggers HA-dependent fusion. The M2 ion channel functions to... 

FIGURE 17.3 Anti-influenza compounds that act as (a) M2 ion channel inhibitors and (b) RNA polymerase inhibitors. 

Amantadine is a symmetrical CIO tricyclic amine with an unusual structure (1-adamantanamine hydrochloride). It interferes with virus uncoating (1) by blocking the M2 ion channel, which is needed to affect a pH change that helps to initiate the uncoating process. Most consistent antiviral activity has been observed against influenza A virus, but amantadine has httle or no activity against influenza B virus (2). However, influenza A virus can become rapidly resistant to amantadine in vitro (3). Amantadine also promotes the release of dopamine from nerve endings, but may also delay its reuptake into synaptic vesicles. 

Rimantadine hydrochloride, an alpha-methyl derivative of amantadine (alpha-methyl-l-adamantane methylamine hydrochloride), is more active than amantadine against influenza A viruses in vitro and in laboratory animals. It is an alternative to amantadine for the prevention and treatment of influenza A virus infections in adults and for the prevention of influenza in children. Adverse effects have been considered to be less common with rimantadine (SEDA-8, 143), and it is generally tolerated better than amantadine, because it causes fewer nervous system adverse effects (1). Unfortunately, rimantadine is more costly, which has led many institutions to develop influenza treatment guidelines. Both drugs work by blocking the M2 ion channel, which is needed to affect a pH change that helps to initiate viral uncoating. 

M. Akeson and D.W. Deamer. Proton conductance by the Gramicidin water wire — model for proton conductance in the Fi Fq ATPases. Biochim. Biophys. Acta, 60 (1990) 101-109. M.S.P. Sansom, I.D. Kerr, and H.S. Son. The influenza A virus M2 ion channel A molecular modeling and simulation study. Virology, 233 (1997) 163-173. 

T. Sakaguchi, L. Quiang, L.H. Pinto, and R.A. Lamb. The active oligomeric state of the minimalistic influenza virus M2 ion channel is a tetramer. Proc. Natl. Acad. Sci. USA, 94 (1997) 5000-5005. 

C. Wang, R.A. Lamb, and L. Pinto. Activation of the M2 ion channel of influenza virus a role for the transmembrane domain histidine residue. Biophys. J., 69 (1995) 1363-1371. 

Stimulation of mAChRs also results in the activation or inhibition of a large number of ion channels [5]. For example, stimulation of Mi receptors leads to the suppression of the so-called M current, a voltage-dependent Recurrent found in various neuronal tissues. M2 receptors, on the other hand, mediate the opening of cardiac Ikcacii) channels, and both M2 and M4 receptors are linked to the inhibition of voltage-sensitive calcium channels [5]. 

The nAChR is comprised of five subunits, each of which spans the lipid bilayer to create a water-filled pore or channel (Fig. la). Each subunit consists of four transmembrane segments, the second transmembrane segment (M2) lines the ion channel (Fig. lb). The extracellular N-terminal domain of every subunit... 

In the resting (closed) state, the ion channel is occluded by a hydrophobic girdle that constitutes a barrier to ion permeation. Agonist binding in the extracellular domain promotes a conformational change that results in a rotational movement of the M2 helices lining the pore, widening the pore by A. This results... 

Pinto EH, Holsinger LJ, Lamb RA (1992) Influenza virus M2 protein has ion channel activity. Cell 69 517-528... 

A G-protein-mediated effect has an absolute requirement for GTP. Reference has already been made to the requirement for GTP in reconstituting hormone-stimulated adenylate cyclase activity. A similar requirement can be demonstrated when the effector is an ion channel, such as the cardiac atrial inward-rectifier K+ channel which is activated following stimulation of the M2 muscarinic acetylcholine receptor. Thus, in the experiment illustrated in Figure 7.8, the channel recorded with a cell-... 

Effects mediated by G-protein coupled receptors (GPCRs) are very much slower than those mediated by, for example, ligand-gated ion channels, primarily because more steps are involved between activation of the receptor and the final response. For example, even in a simple, three-step, G-protein-mediated effect, such as the opening of atrial GIRK channels following the activation of M2 muscarinic receptors by acetylcholine, which follows the scheme ... 

Leonard, R.J., Labarca, C.G., Chamet, P., Davidson, N., Lester, H.A. Evidence that the M2 membrane-spanning region lines the ion channel pore of the nicotinic receptor. Science. 242 1578, 1988. 

Currently, two classes of drugs are available with antiviral activity against influenza viruses inhibitors of the ion channel activity of the M2 membrane protein, amantadine and rimantadine, and the neuraminidase inhibitors oseltamivir, and zanamivir. H5N1 viruses isolated from poultry and humans in Thailand and Viet Nam in 2004 invariably showed an amantadine-resistance indicating that amantadine treatment is not an option during the ongoing outb-treak in South-East Asia. 

Ion channels. G proteins are also involved in the coupling of some receptors to ion channels. A classic example is the modulation of inwardly rectifying potassium channels by m2 acetylcholine receptors. Other receptors have been shown to modulate Ca and/or K channels by either G protein a or Py subunits. 

Fig. 16.12. Subunit structure of the acetylcholine receptor, a) The acetylcholine receptor has the subunit structure 02 7 - The four transmembrane elements Ml—M4 are shown for the y subunit. The binding sites for acetylcholine (ACh) are located on the a-subunits. b) It is assumed that the inner wall of the ion pore is formed by M2 helices of the five subunits, c) Postulated configuration of the M2 helices in the narrowest region of the ion channel. In the closed state, five leucine residues (one per subunit) lie in the ion channel and hinder passage of ions. Above and below the block, there are negatively charged residues that serve as prefilters for ion passage.

Fig. 16.13. Pore structure of the acetylcholine receptor, based on electron microscopy studies. a) Electron density map of the acetylcholine receptor of the postsynaptic membrane of the electric organ of the ray Torpedo californicus, based on electron microscopy studies. The receptor has a long funnel-like structure in the extracellular region, which narrows at the center of the pore. A smaller funnel structure is observed in the cytoplasmic region of the receptor. Another protein is situated on the cytoplasmic side. The long arrow indicates the direction of ion passage and the small arrow shows the postulated binding site for acetylcholine, b) Schematic representation of the acetylcholine receptor with the M2 hehx as the central block in the ion channel. According to Unwin, (1993).

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