Functional characterization of amino acids that could be involved in zinc(II)-binding.

For cell surface expression studies COS-7 cells were transiently transfected with the empty expression vector pcDps (mock), Gpr83 wild type or Gpr83 mutants. HEK293 cells were used for functional characterization. Data were evaluated from three or four independent experiments, each performed at least in triplicates. IP3 accumulation performed as reporter gene assay was calculated fold over the basal mock transfection with 24718.3 ± 3958.7 relative light units, set to 1. The hTSHR stimulated with 100 mU/ml bTSH functioned as control for Gq/11 activation [data not shown, [31], [32]]. Shown data represent mean ± SEM. The mutated aminoacid residues are grouped into extracellular located Ds (Asp) and És (Glu), Hs (His) and one C (Cys) that could be involved in Zn(II)-binding. Asteriks indicate significant higher basal activity in comparison to wild type. ** p