Signaling states of rhodopsin. Formation of the storage form, metarhodopsin III, from active metarhodopsin II.
Vertebrate rhodopsin consists of the apoprotein opsin and the chromophore 11-cis-retinal covalently linked via a protonated Schiff base. Upon photoisomerization of the chromophore to all-trans-retinal, the retinylidene linkage hydrolyzes, and all-trans-retinal dissociates from opsin. The pigment is eventually restored by recombining with enzymatically produced 11-cis-retinal. All-trans-retinal release occurs in parallel with decay of the active form, metarhodopsin (Meta) II, in which the original Schiff base is intact but deprotonated. The intermediates formed during Meta II decay include Meta III, with the original Schiff base reprotonated, and Meta III-like pseudo-photoproducts. Using an intrinsic fluorescence assay, Fourier transform infrared spectroscopy, and UV-visible spectroscopy, we investigated Meta II decay in native rod disk membranes. Up to 40% of Meta III is formed without changes in the intrinsic Trp fluorescence and thus without all-trans-retinal release. NADPH, a cofactor for the reduction of all-trans-retinal to all-trans-retinol, does not accelerate Meta II decay nor does it change the amount of Meta III formed. However, Meta III can be photoconverted back to the Meta II signaling state. The data are described by two quasi-irreversible pathways, leading in parallel into Meta III or into release of all-trans-retinal. Therefore, Meta III could be a form of rhodopsin that is stored away, thus regulating photoreceptor regeneration.
Heck
Martin
M
Institut für Medizinische Physik und Biophysik, Universitätsklinikum Charité, Humboldt Universität zu Berlin, Schumannstrasse 20-21, 10098 Berlin, Germany. martin.heck@charite.de
Schädel
Sandra A
SA
Maretzki
Dieter
D
Bartl
Franz J
FJ
Ritter
Eglof
E
Palczewski
Krzysztof
K
Hofmann
Klaus Peter
KP
D000818:Animals; D002417:Cattle; D007536:Isomerism; D007700:Kinetics; D008027:Light; D008958:Models, Molecular; D010777:Photochemistry; D017433:Protein Structure, Secondary; D017948:Retinal Rod Photoreceptor Cells / Q000502:physiology*; D012172:Retinaldehyde / Q000378:metabolism; D012243:Rhodopsin / Q000031:analogs & derivatives* / Q000502:physiology*; D017299:Rod Opsins / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D013053:Spectrophotometry; D017550:Spectroscopy, Fourier Transform Infrared
D017299:Rod Opsins; C013110:metarhodopsins; D012243:Rhodopsin; D012172:Retinaldehyde