The gecko visual pigment: its photosensitivity and the effects of chloride and nitrate ions

By use of the method of photometric curves, the photosensitivity of the major and ion-sensitive pigment of Gekko gekko has been determined and compared with that of rhodopsins of the frog (Rana pipiens) and of the fish (Porichthys notatus). In the presence of Cl- (or Br-), the gecko pigment has the same photosensitivity as the other A1 rod pigments, but unlike these, the addition of NH2OH does not lead to a Dartnall effect, i.e. an enhancement in the measured rate of photic bleaching. This is because the gecko pigment has no meta-III intermediate. In the Cl- -deficient state the gecko pigment has a photosensitivity 0.8 times that of the Cl- -provided system. The increase in photosensitivity brought on by Cl- is quantitatively accounted for by the Cl- -induced hyperchromic effect. The addition of NH2OH to the system without added Cl- leads to a small increase in measured rate of photic bleaching with an apparent 13% increment in photosensitivity. This is not a classical Dartnall effect for here again no meta-III is involved. The possibility is raised of an additional, yet undiscovered, action of NH2OH on the opsin moiety. Nitrate ions (NO3-) are known to produce an increase in extinction coefficient similar to that of Cl- and a hypochromic shift in the spectral absorbance. Despite the hyperchromic action, NO3- produces a reduction in the measured rate of photic bleaching, an effect explained by the appearance of a meta-III type intermediate absorbing at about 470 nm. While Cl- is able to antagonize the NO3- -induced hypochromic shift, it is unable to reverse the NO3- -induction of meta-III. This, along with other differences in responses of the gecko pigment to these two ions, suggests that Cl- and NO3- act at two different sites and produce unique conformational changes in the protein molecule.