A Quantitative Model of the GIRK1/2 Channel Reveals That Its Basal and Evoked Activities Are Controlled by Unequal Stoichiometry of G? and G??.

ABSTRACT: G protein-gated K+ channels (GIRK; Kir3), activated by G?? subunits derived from Gi/o proteins, regulate heartbeat and neuronal excitability and plasticity. Both neurotransmitter-evoked (Ievoked) and neurotransmitter-independent basal (Ibasal) GIRK activities are physiologically important, but mechanisms of Ibasal and its relation to Ievoked are unclear. We have previously shown for heterologously expressed neuronal GIRK1/2, and now show for native GIRK in hippocampal neurons, that Ibasal and Ievoked are interrelated: the extent of activation by neurotransmitter (activation index, Ra) is inversely related to Ibasal. To unveil the underlying mechanisms, we have developed a quantitative model of GIRK1/2 function. We characterized single-channel and macroscopic GIRK1/2 currents, and surface densities of GIRK1/2 and G?? expressed in Xenopus oocytes. Based on experimental results, we constructed a mathematical model of GIRK1/2 activity under steady-state conditions before and after activation by neurotransmitter. Our model accurately recapitulates Ibasal and Ievoked in Xenopus oocytes, HEK293 cells and hippocampal neurons; correctly predicts the dose-dependent activation of GIRK1/2 by coexpressed G?? and fully accounts for the inverse Ibasal-Ra correlation. Modeling indicates that, under all conditions and at different channel expression levels, between 3 and 4 G?? dimers are available for each GIRK1/2 channel. In contrast, available G?i/o decreases from ~2 to less than one G? per channel as GIRK1/2's density increases. The persistent G??/channel (but not G?/channel) ratio support a strong association of GIRK1/2 with G??, consistent with recruitment to the cell surface of G??, but not G?, by GIRK1/2. Our analysis suggests a maximal stoichiometry of 4 G?? but only 2 G?i/o per one GIRK1/2 channel. The unique, unequal association of GIRK1/2 with G protein subunits, and the cooperative nature of GIRK gating by G??, underlie the complex pattern of basal and agonist-evoked activities and allow GIRK1/2 to act as a sensitive bidirectional detector of both G?? and G?.

SUBMITTER: Yakubovich D 

PROVIDER: S-EPMC4636287 | biostudies-literature | 2015 Nov

REPOSITORIES: biostudies-literature