Chloride binding regulates the schiff base pK in gecko P521 cone-type visual pigment

Ching Yuan, Osamu Kuwata, Jie Liang, Saurav Misra, Sergei P. Balashov, Thomas G. Ebrey

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

The binding of chloride is known to shift the absorption spectrum of most long-wavelength-absorbing cone-type visual pigments roughly 30 nm to the red. We determined that the chloride binding constant for this color shift in the gecko P521 visual pigment is 0.4 mM at pH 6.0. We found an additional effect of chloride on the P521 pigment: the apparent pK(a) of the Schiff base in P521 is greatly increased as the chloride concentration is increased. The apparent Schiff base pK(a) shifts from 8.4 for the chloride-free form to >10.4 for the chloride-bound form. We show that this shift is due to chloride binding to the pigment, not to the screening of the membrane surface charges by chloride ions. We also found that at high pH, the absorption maximum of the chloride-free pigment shifts from 495 to 475 nm. We suggest that the chloride-dependent shift of the apparent Schiff base pK(a) is due to the deprotonation of a residue in the chloride binding site with a pK(a) of ca. 8.5, roughly that of the Schiff base in the absence of chloride. The deprotonation of this site results in the formation of the 475 nm pigment and a 100-fold decrease in the pigment's ability to bind chloride. Increasing the concentration of chloride results in the stabilization of the protonated state of this residue in the chloride binding site and thus increased chloride binding with an accompanying increase in the Schiff base pK.

Original languageEnglish (US)
Pages (from-to)4649-4654
Number of pages6
JournalBiochemistry
Volume38
Issue number14
DOIs
StatePublished - Apr 6 1999

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