期刊
ACS CHEMICAL NEUROSCIENCE
卷 12, 期 3, 页码 562-572出版社
AMER CHEMICAL SOC
DOI: 10.1021/acschemneuro.0c00781
关键词
Structure- unction relationship; electrostatic interaction; Cys-Loop receptor; gating; histidine; patch clamp
资金
- Polish National Science Centre grant MAESTRO [DEC-2015/18/A/NZ1/00395]
The GABA(A) receptor, a member of the Cys-loop family, plays a crucial role in brain inhibition, but the molecular mechanisms underlying its conformational transitions remain uncertain. In this study, mutations of H55 were found to primarily affect preactivation and desensitization transitions of the receptor, likely through influencing local electrostatic interactions at the receptor interface. Positive and negative residue substitutions at H55 resulted in opposite effects on receptor gating, indicating the involvement of electrostatic interactions in these processes.
The GABA(A) receptor is a member of the Cys-loop family and plays a crucial role in the adult mammalian brain inhibition. Although the static structure of this receptor is emerging, the molecular mechanisms underlying its conformational transitions remain elusive. It is known that in the Cys-loop receptors, the interface between extracellular and transmembrane domains plays a key role in transmitting the activation wave down to the channel gate in the pore. It has been previously reported that histidine 55 (H55), located centrally at the interfacial beta 1-beta 2 loop of the alpha(1) subunit, is important in the receptor activation, but it is unknown which specific gating steps it is affecting. In the present study, we addressed this issue by taking advantage of the state-of-the-art macroscopic and single-channel recordings together with extensive modeling. Considering that H55 is known to affect the local electrostatic landscape and because it is neighbored by two negatively charged aspartates, a well conserved feature in the alpha subunits, we considered substitution with negative (E) and positive (K) residues. We found that these mutations markedly affected the receptor gating, altering primarily preactivation and desensitization transitions. Importantly, opposite effects were observed for these two mutations strongly suggesting involvement of electrostatic interactions. Single-channel recordings suggested also a minor effect on opening/closing transitions which did not depend on the electric charge of the substituting amino acid. Altogether, we demonstrate that H55 mutations affect primarily preactivation and desensitization most likely by influencing local electrostatic interactions at the receptor interface.
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