Figure 1
From: Calmodulin interacts with Rab3D and modulates osteoclastic bone resorption
Calmodulin interacts with Rab3D.
(A) A yeast two hybrid assay showing that Calmodulin interacts with Rab3D, by using histidine-deficient plate. (B) BRET assays showing that co-transfection of Rluc-Camodulin and EYFP-Rab3D fusion protein constructs resulted in a significant BRET signal. Co-expression of Rluc and EYFP is shown as a negative control. (C) Flag-Rab3D proteins expressed in COS cells interact with calmodulin saphorose in the presence of 2 mM calcium. *Indicates p Value < 0.001 when compared with EYFP and Rluc. (D) Calmodulin calcium-insensitive mutant perturbs its interaction with Rab3D. Generation of a Rluc-calmodulin construct in which four aspartic acid residues at position 23, 59, 96, 132 were substituted with alanine, mimicking a calcium insensitive form of calmodulin. (E) BRET assays showing that the calcium insensitive form of camodulin failed to interact with Rab3D. 1:1, 1:2 and 1:3 indicate that transfected plasmid ratio of EYFP-Rab3D/ Rluc-camodulin or EYFP-Rab3D/ Rluc-calmodulin mutant 1234. Symbol *indicates p Value < 0.001 when compared with EYFP and Rluc-camodulin control. Symbol # indicates p Value < 0.001 when compared Rluc-camodulin with Rluc-calmodulin mutant 1234.
