Multiple tyrosine residues at the GABA binding pocket influence surface expression and mediate kinetics of the GABA(A) receptor

作者：Laha Kurt T^*; Tran Phu N

来源：Journal of Neurochemistry, 2013, 124(2): 200-209.

DOI：10.1111/jnc.12083

• GABA binding
• GABA(A) receptor
• kinetics
• surface expression

The prevalence of aromatic residues in the ligand binding site of the GABA(A) receptor, as with other cys-loop ligand-gated ion channels, is undoubtedly important for the ability of neurotransmitters to bind and trigger channel opening. Here, we have examined three conserved tyrosine residues at the GABA binding pocket (beta(2)Tyr97, beta(2)Tyr157, and beta(2)Tyr205), making mutations to alanine and phenylalanine. We fully characterized the effects each mutation had on receptor function using heterologous expression in HEK-293 cells, which included examining surface expression, kinetics of macroscopic currents, microscopic binding and unbinding rates for an antagonist, and microscopic binding rates for an agonist. The assembly or trafficking of GABA(A) receptors was disrupted when tyrosine mutants were expressed as alpha beta receptors, but interestingly not when expressed as alpha beta gamma receptors. Mutation of each tyrosine accelerated deactivation and slowed GABA binding. This provides strong evidence that these residues influence the binding of GABA. Qualitatively, mutation of each tyrosine has a very similar effect on receptor function; however, mutations at beta(2)Tyr157 and beta(2)Tyr205 are more detrimental than beta(2)Tyr97 mutations, particularly to the GABA binding rate. Overall, the results suggest that interactions involving multiple tyrosine residues are likely during the binding process.

• 出版日期2013-1