Calcium-dependent binding of calmodulin to neuronal gap junction proteins.
We examined the interactions of calmodulin with neuronal gap junction proteins connexin35 (Cx35) from perch, its mouse homologue Cx36, and the related perch Cx34.7 using surface plasmon resonance. Calmodulin bound to the C-terminal domains of all three connexins with rapid kinetics in a concentration- and Ca2+-dependent manner. Dissociation was also very rapid. K(d)'s for calmodulin binding at a high-affinity site ranged from 11 to 72 nM, and K(1/2)'s for Ca2+ were between 3 and 5 microM. No binding to the intracellular loops was observed. Binding competition experiments with synthetic peptides mapped the calmodulin binding site to a 10-30 amino acid segment at the beginning of the C-terminal domain of Cx36. The micromolar K(1/2)'s and rapid on and off rates suggest that this interaction may change dynamically in neurons, and may occur transiently when Ca2+ is elevated to a level that would occur in the near vicinity of an activated synapse.
Burr
Gary S
GS
Department of Ophthalmology and Visual Science, University of Texas Health Science Center at Houston, USA.
Mitchell
Cheryl K
CK
Keflemariam
Yenabi J
YJ
Heidelberger
Ruth
R
O'Brien
John
J
D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000737:chemistry*; D002147:Calmodulin / Q000737:chemistry*; D017630:Connexins / Q000737:chemistry*; D005136:Eye Proteins / Q000737:chemistry*; D029941:Fish Proteins / Q000737:chemistry*; D051379:Mice; D009474:Neurons / Q000737:chemistry*; D010471:Perches; D011485:Protein Binding; D000097004:Gap Junction delta-2 Protein
D002118:Calcium; D002147:Calmodulin; D017630:Connexins; D005136:Eye Proteins; D029941:Fish Proteins; D000097004:Gap Junction delta-2 Protein; C114667:connexin 34.7 protein, Morone americana; C100585:connexin 35 protein, vertebrate