Resonance raman spectroscopy of an ultraviolet-sensitive insect rhodopsin.
We present the first visual pigment resonance Raman spectra from the UV-sensitive eyes of an insect, Ascalaphus macaronius (owlfly). This pigment contains 11-cis-retinal as the chromophore. Raman data have been obtained for the acid metarhodopsin at 10 degrees C in both H2O and D2O. The C = N stretching mode at 1660 cm-1 in H2O shifts to 1631 cm-1 upon deuteriation of the sample, clearly showing a protonated Schiff base linkage between the chromophore and the protein. The structure-sensitive fingerprint region shows similarities to the all-trans-protonated Schiff base of model retinal chromophores, as well as to the octopus acid metarhodopsin and bovine metarhodopsin I. Although spectra measured at -100 degrees C with 406.7-nm excitation, to enhance scattering from rhodopsin (lambda max 345 nm), contain a significant contribution from a small amount of contaminants [cytochrome(s) and/or accessory pigment] in the sample, the C = N stretch at 1664 cm-1 suggests a protonated Schiff base linkage between the chromophore and the protein in rhodopsin as well. For comparison, this mode also appears at approximately 1660 cm-1 in both the vertebrate (bovine) and the invertebrate (octopus) rhodopsins. These data are particularly interesting since the absorption maximum of 345 nm for rhodopsin might be expected to originate from an unprotonated Schiff base linkage. That the Schiff base linkage in the owlfly rhodopsin, like in bovine and in octopus, is protonated suggests that a charged chromophore is essential to visual transduction.
Physics Department, City College of New York, New York 10031.
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Callender
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Schwemer
D000818:Animals; D003624:Darkness; D004175:Diptera; D008027:Light; D012160:Retina / Q000378:metabolism; D012168:Retinal Pigments / Q000528:radiation effects*; D012243:Rhodopsin / Q000031:analogs & derivatives / Q000378:metabolism / Q000528:radiation effects*; D013059:Spectrum Analysis, Raman / Q000379:methods; D014466:Ultraviolet Rays
D012168:Retinal Pigments; C013110:metarhodopsins; D012243:Rhodopsin