Human nACh a4 -- +Leu 260

Species Insertion Mutation
Human

Leu 260

+L260
(L776ins3)

Rat Equivalent Leu 264

+L264
(L776ins3)

Mouse Equivalent    

h +Leu 260

The apparent affinity for ACh of this mutant was found to be 0.28 mM, compared to 2mM for the control. There was a small shift in the desensitization IC50.

EC50 ACh (mM) nH IC50 ACh (mM) nH EC50 cytisine (mM) nH IMac ACh (nA)
ha4(776ins3)b2 .34 +/- 0.2 0.92 +/- 0.20 0.038 +/- 0.016 1.2 +/- 0.11 0.042 +/- 0.02 0.67 +/- 0.08 1624 +/- 569
control

3.4 +/- 2.7

0.70 +/- 0.05 0.11 +/- 0.02 1.4 +/- 0.15 2.12 +/- 1.7 0.79 +/- 0.14 1762 +/- 439

 

Epibatidine
nH
Epibatidine
Log intercept
Acetylcholine
nH
Acetylcholine
Log intercept
Nictoine
nH
Nicotine
Log intercept
ha4(776ins3)b2 1.12 +/- 0.13 -5.63 +/- 0.4 1.15 +/- 0.14 -3.55 +/- 0.36 1.12 +/- 0.22 -3.62 +/- 0.64
control 1.21 +/- 0.17 -5.13 +/- 0.32 1.2 +/- 0.27 -3.16 +/- 0.47 1.24 +/- 0.27 -3.10 +/- 0.62

In the control receptor cytisine evoked about 24.77% of ACh currents, but with this mutation it evoked about 78.2%. The EC50 for cytisine in the control was 2.12 mM, but in the mutant receptors, it was 0.042 mM.

Bertrand S, Weiland S, Berkovic SF, Steinlein OK, Bertrand D (1998) Properties of Neuronal Nictoinic Acetylcholine Receptor Mutants from Humans Suffering from Autosomal Dominant Nocturnal Frontal Lobe Epilepsy. British Journal of Pharmacology 125:751-760

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Incubation with 320 mM Carbamazepine inhibited the ACh-evoked currents. The effect on the V287M mutant was greater than on the S252L mutant. a4S248F, a4L776ins3 and b2V287M are all more sensitive to CBZ than a4S252L.

Bertrand D, Picard F, Hellard SL, Weiland S, Favre I, Phillips H, Bertrand S, Berkovic SF, Malafosse A, Mulley J (2002) How Mutations in the nAChRs Can Cause ADNFLE Epilepsy. Epilepsia 43(Suppl. 5):112-122

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