We have quantitatively investigated specificities in activating G-protein subtype by bovine and squid rhodopsins to examine whether or not the phototransduction cascade in each of the photoreceptor cells is determined by the colocalization of a large amount of G-protein subtype (Gt or Gq). In contrast to the efficient activation of respective Gt and Gq, bovine and squid rhodopsins scarcely activated G-protein counterparts. Exchange of alpha- and betagamma-subunits of Gt and Gq indicated the critical role of the alpha-subunit in specific binding to respective rhodopsins. Thus the specific recognition of G-protein subtype by each rhodopsin is a major mechanism in determining the phototransduction cascade.