Cloning and in silico investigation of a putative voltage-gated calcium channel gene and protein in Astacus Leptodactylus

Objectives: Voltage-gated calcium channels are essential elements in development of many cellular processes like electrical signaling, contraction secretion and gene expression. There has been a fair amount of information about the functional and structural properties of the calcium channels in mammalian species. Crayfish serves as a model animal for many types of experiments. However, there has been no information related to the molecular and genetic properties of the calcium channels in the crayfish.Methods: Conventional cloning methods, three-dimensional structural calculations, docking experiments have been conducted.Results: An mRNA 7,791 bp in size has been cloned. The coding region has been translated into an alpha peptide with 1,942 residues. The cloned protein sequence has similarity to other L-type voltage-gated calcium channel sequences from the neighboring species. Three-dimensional structure, in reference to human L-type voltage-gated calcium channel, has been calculated. Known calcium channel blockers, nifedipine, verapamil and diltiazem have been successfully docked on the calculated three-dimensional model.Conclusions: Considering the similarity assay in the National Center for Biotechnology Information (NCBI) platform, the three-dimensional structural calculations and the docking experiments it was concluded that the cloned mRNA codes an alpha peptide for a putative voltage-gated calcium channel protein in the crayfish. In the present work by using the conventional molecular biology methods a complete mRNA coding a putative calcium channel has been de novo cloned. Three-dimensional structure of the related protein has been calculated and several pharmacological agents blocking the channel have been docked to the identified receptor sites.