Beta subunit phosphorylation selectively increases fast desensitization and prolongs deactivation of alpha1beta1gamma2L and alpha1beta3gamma2L GABA(A) receptor currents.
We studied the effects of phosphorylation by protein kinase A (PKA) on GABA(A) receptors (alpha1beta1gamma2L andalpha1beta3gamma2L) transiently expressed in HEK 293T cells. Under conditions favorable for PKA activation, currents obtained using whole-cell patch clamp of lifted cells displayed increased rate and extent of the fast phases of desensitization, decreased rate of current deactivation after GABA removal, and prolongation of brief IPSC-like currents. Mutation of serine residues (beta1 S409, beta3 S407, beta3 S408) revealed that only beta1 S409 and beta3 S408 were critical for the modulatory effect of PKA on GABA(A) receptor currents. Additionally, repeated pulse inhibition was increased in receptors after mutation of the critical serine to glutamate and decreased when the serine was mutated to alanine. These data demonstrate that PKA phosphorylation modulated GABA(A) receptor currents by increasing fast phases of macroscopic desensitization and suggest a role for PKA in regulating GABAergic IPSC duration.
Hinkle
David J
DJ
Neuroscience Graduate Program, University of Michigan, Ann Arbor, Michigan 48104-1687, USA.
Macdonald
Robert L
RL
D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D016895:Culture Media, Serum-Free; D000242:Cyclic AMP / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004553:Electric Conductivity; D006801:Humans; D007700:Kinetics; D009154:Mutation; D018408:Patch-Clamp Techniques; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D010766:Phosphorylation; D021122:Protein Subunits; D011963:Receptors, GABA-A / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012694:Serine / Q000378:metabolism; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
D016895:Culture Media, Serum-Free; C546240:GABRA1 protein, human; C546273:GABRB3 protein, human; C546279:GABRG2 protein, human; D021122:Protein Subunits; D011963:Receptors, GABA-A; D005576:Colforsin; D012694:Serine; D005680:gamma-Aminobutyric Acid; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D010749:Phosphoprotein Phosphatases