The pKa of the protonated Schiff bases of gecko cone and octopus visual pigments.
A visual pigment is composed of retinal bound to its apoprotein by a protonated Schiff base linkage. Light isomerizes the chromophore and eventually causes the deprotonation of this Schiff base linkage at the meta II stage of the bleaching cycle. The meta II intermediate of the visual pigment is the active form of the pigment that binds to and activates the G protein transducin, starting the visual cascade. The deprotonation of the Schiff base is mandatory for the formation of meta II intermediate. We studied the proton binding affinity, pKa, of the Schiff base of both octopus rhodopsin and the gecko cone pigment P521 by spectral titration. Several fluorinated retinal analogs have strong electron withdrawing character around the Schiff base region and lower the Schiff base pKa in model compounds. We regenerated octopus and gecko visual pigments with these fluorinated and other retinal analogs. Experiments on these artificial pigments showed that the spectral changes seen upon raising the pH indeed reflected the pKa of the Schiff base and not the denaturation of the pigment or the deprotonation of some other group in the pigment. The Schiff base pKa is 10.4 for octopus rhodopsin and 9.9 for the gecko cone pigment. We also showed that although the removal of Cl- ions causes considerable blue-shift in the gecko cone pigment P521, it affects the Schiff base pKa very little, indicating that the lambda max of visual pigment and its Schiff base pKa are not tightly coupled.
Liang
J
J
Biophysics Program, University of Illinois at Urbana-Champaign 61801.
Steinberg
G
G
Livnah
N
N
Sheves
M
M
Ebrey
T G
TG
Tsuda
M
M
D000818:Animals; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008027:Light; D008116:Lizards; D008871:Microvilli / Q000502:physiology; D015394:Molecular Structure; D049831:Octopodiformes; D017949:Retinal Cone Photoreceptor Cells / Q000502:physiology*; D012168:Retinal Pigments / Q000737:chemistry* / Q000378:metabolism; D012172:Retinaldehyde / Q000737:chemistry* / Q000378:metabolism; D012545:Schiff Bases; D013053:Spectrophotometry
D012168:Retinal Pigments; D012545:Schiff Bases; D012172:Retinaldehyde