Difference in molecular structure of rod and cone visual pigments studied by Fourier transform infrared spectroscopy.
To investigate the local structure that causes the differences in molecular properties between rod and cone visual pigments, we have measured the difference infrared spectra between chicken green and its photoproduct at 77 K and compared them with those from bovine and chicken rhodopsins. In contrast to the similarity of the vibrational bands of the chromophore, those of the protein part were notably different between chicken green and the rhodopsins. Like the rhodopsins, chicken green has an aspartic acid at position 83 (D83) but exhibited no signals due to the protonated carboxyl of D83 in the C=O stretching region, suggesting that the molecular contact between D83 and G120 through water molecule evidenced in bovine rhodopsin is absent in chicken green. A pair of positive and negative bands due to the peptide backbone (amide I) was prominent in chicken green, while the rhodopsins exhibited only small bands in this region. Furthermore, chicken green exhibited characteristic paired bands around 1480 cm(-1), which were identified as the imide bands of P189 using site-directed mutagenesis. P189, situated in the putative second extracellular loop, is conserved in all the known cone visual pigments but not in rhodopsins. Thus, some region of the second extracellular loop including P189 is situated near the chromophore and changes its environment upon formation of the batho-intermediate. The results noted above indicate that differences in the protein parts between chicken green and the rhodopsins alter the changes seen in the protein upon photoisomerization of the chromophore. Some of these changes appear to be the pathway from the chromophore to cytoplasmic surface of the pigment and thus could affect the activation process of transducin.
Department of Biophysics, Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan.
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H
Imai
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Hirano
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H
Kandori
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A
Terakita
Y
Y
Shichida
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D030161:Avian Proteins; D002417:Cattle; D002645:Chickens; D005136:Eye Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007094:Imides / Q000032:analysis; D008027:Light; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D020419:Photoreceptor Cells, Vertebrate / Q000737:chemistry* / Q000378:metabolism; D011392:Proline / Q000737:chemistry / Q000235:genetics; D017433:Protein Structure, Secondary; D012168:Retinal Pigments / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012243:Rhodopsin / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods
D030161:Avian Proteins; D005136:Eye Proteins; D007094:Imides; D012168:Retinal Pigments; C088721:RHO rhodopsin protein, chicken; D012243:Rhodopsin; D011392:Proline