Dynamic aspects of functional regulation of the ATP receptor channel P2X2.
The P2X(2) channel is a ligand-gated channel activated by ATP. Functional features that reflect the dynamic flexibility of the channel include time-dependent pore dilatation following ATP application and direct inhibitory interaction with activated nicotinic acetylcholine receptors on the membrane. We have been studying the mechanisms by which P2X(2) channel functionality is dynamically regulated. Using a Xenopus oocyte expression system, we observed that the pore properties, including ion selectivity and rectification, depend on the open channel density on the membrane. Pore dilatation was apparent when the open channel density was high and inward rectification was modest. We also observed that P2X(2) channels show voltage dependence, despite the absence of a canonical voltage sensor. At a semi-steady state after ATP application, P2X(2) channels were activated upon membrane hyperpolarization. This voltage-dependent activation was also [ATP] dependent. With increases in [ATP], the speed of hyperpolarization-induced activation was increased and the conductance-voltage relationship was shifted towards depolarized potentials. Based on analyses of experimental data and various simulations, we propose that these phenomena can be explained by assuming a fast ATP binding step and a rate-limiting voltage-dependent gating step. Complete elucidation of these regulatory mechanisms awaits dynamic imaging of functioning P2X(2) channels.
Kubo
Yoshihiro
Y
Division of Biophysics and Neurobiology, Department of Molecular Physiology, National Institute for Physiological Sciences, Okazaki, Aichi, Japan. ykubo@nips.ac.jp
Fujiwara
Yuichiro
Y
Keceli
Batu
B
Nakajo
Koichi
K
D000595:Amino Acid Sequence; D000818:Animals; D006801:Humans; D056004:Molecular Dynamics Simulation / Q000639:trends*; D008969:Molecular Sequence Data; D018048:Receptors, Purinergic P2 / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology*; D058476:Receptors, Purinergic P2X2; D013569:Synapses / Q000737:chemistry / Q000378:metabolism / Q000502:physiology
C546401:P2RX2 protein, human; D018048:Receptors, Purinergic P2; D058476:Receptors, Purinergic P2X2