Purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina

Fukagawa, Takashi; Takafuji, Kazuaki; Tachibanaki, Shuji; Kawamura, Sadao

doi:10.1371/journal.pone.0173908

Cited by 7 publications

(6 citation statements)

References 49 publications

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“…5A). The preparation of the visual pigments from carp fish were performed as described previously in (59). The detergent extracted carp fish pigments, mostly had rods in it, but also ~5% cones; this was determined by radiating the sample with deep red light and measuring the difference absorption spectra which showed the presence of red cones (see Supporting Information for more details, Figure S3).…”

Section: Methodsmentioning

confidence: 99%

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Comparison of Bovine and Carp Fish Visual Pigment Photo‐Intermediates at Room Temperature

Dhankhar

Nagpal

Tachibanaki

et al. 2022

Photochem & Photobiology

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This paper presents room temperature nanoseconds to milliseconds time‐resolved spectra and kinetics of the intermediate states and species of bovine and carp fish rhodopsin visual pigments, which also contained ~5% cone pigments. The nanoseconds to milliseconds range cover all the major intermediates in the visual phototransduction process except the formation of bathorhodopsin intermediate which occurs at the femtosecond time scale. The dynamics of these visual pigment intermediates are initiated by excitation with a 532 nm nanosecond laser pulse. The recorded differences between bovine and carp rhodopsin time‐resolved spectra of the formation and decay kinetics of their intermediates are presented and discussed. The data show that the carp samples batho intermediate decays faster, nearly by a factor of three, compared to the bovine samples. The formation and decay spectra and kinetics of rhodopsin outer segments and extracted rhodopsin inserted in buffer solution were found to be identical, with very small differences between them in the decay lifetimes of bathorhodopsin and formation of lumirhodopsin.

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Section: Methodsmentioning

confidence: 99%

“…The rhodopsin protein was extracted using the procedure described by Tachibanaki et al . which involved a detergent in a buffer (57–59) that consisted of 20% w/v glycerol, 2 mM MgCl 2 , 140 mM NaCl, 50 mM Hepes and 1 mM DTT. ROS were added to ~5 mL buffer and 50 mg DMM detergent.…”

Section: Methodsmentioning

confidence: 99%

Comparison of Bovine and Carp Fish Visual Pigment Photo‐Intermediates at Room Temperature

Dhankhar

Nagpal

Tachibanaki

et al. 2022

Photochem & Photobiology

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“…The visual pigments, rhodopsin (Rh) in rods and three types of pigments in three types of human cones (S-, M-, and L-opsin), are also the main proteins present in the photoreceptor outer segments [ 12 , 13 , 14 , 15 ]. Such a high density of visual pigments, which reaches up to 20,000 molecules per μm 2 of disc membrane in the case of Rh [ 16 ], allows effective absorption of light.…”

Section: Introductionmentioning

confidence: 99%

Structural Impact of Selected Retinoids on Model Photoreceptor Membranes

Radzin,

Wiśniewska-Becker,

Markiewicz

et al. 2023

Membranes

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Photoreceptor membranes have a unique lipid composition. They contain a high level of polyunsaturated fatty acids including the most unsaturated fatty acid in nature, docosahexaenoic acid (22:6), and are enriched in phosphatidylethanolamines. The phospholipid composition and cholesterol content of the subcellular components of photoreceptor outer segments enables to divide photoreceptor membranes into three types: plasma membranes, young disc membranes, and old disc membranes. A high degree of lipid unsaturation, extended exposure to intensive irradiation, and high respiratory demands make these membranes sensitive to oxidative stress and lipid peroxidation. Moreover, all-trans retinal (AtRAL), which is a photoreactive product of visual pigment bleaching, accumulates transiently inside these membranes, where its concentration may reach a phototoxic level. An elevated concentration of AtRAL leads to accelerated formation and accumulation of bisretinoid condensation products such as A2E or AtRAL dimers. However, a possible structural impact of these retinoids on the photoreceptor-membrane properties has not yet been studied. In this work we focused just on this aspect. The changes induced by retinoids, although noticeable, seem not to be significant enough to be physiologically relevant. This is, however, an positive conclusion because it can be assumed that accumulation of AtRAL in photoreceptor membranes will not affect the transduction of visual signals and will not disturb the interaction of proteins engaged in this process.

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“…Few model animals have a cone-dominated retina; ground squirrel and tree shrew are notable exceptions [ 11 , 12 ]. As a consequence, the preparation of relatively pure cone outer segments suitable for biochemical characterization of transduction components is often not possible although progress in cone purification techniques has been made, for example, with carp [ 13 , 14 ]. While the general structure of the signaling cascade and its shutoff mechanisms are similar in rods and cones, cones use many distinct phototransduction proteins including critical components of the cascade: photopigment, G-protein transducin, effector phosphodiesterase, cyclic nucleotide gated channel (reviewed in [ 15 ]).…”

Section: Introductionmentioning

confidence: 99%

Phototransduction in retinal cones: Analysis of parameter importance

et al. 2021

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In daylight, cone photoreceptors in the retina are responsible for the bulk of visual perception, yet compared to rods, far less is known quantitatively about their biochemistry. This is partly because it is hard to isolate and purify cone proteins. The issue is also complicated by the synergistic interaction of these parameters in producing systems biology outputs, such as photoresponse. Using a 3-D resolved, finite element model of cone outer segments, here we conducted a study of parameter significance using global sensitivity analysis, by Sobol indices, which was contextualized within the uncertainty surrounding these parameters in the available literature. The analysis showed that a subset of the parameters influencing the circulating dark current, such as the turnover rate of cGMP in the dark, may be most influential for variance with experimental flash response, while the shut-off rates of photoexcited rhodopsin and phosphodiesterase also exerted sizable effect. The activation rate of transducin by rhodopsin and the light-induced hydrolysis rate of cGMP exerted measurable effects as well but were estimated as relatively less significant. The results of this study depend on experimental ranges currently described in the literature and should be revised as these become better established. To that end, these findings may be used to prioritize parameters for measurement in future investigations.

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Purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina

Cited by 7 publications

References 49 publications

Comparison of Bovine and Carp Fish Visual Pigment Photo‐Intermediates at Room Temperature

Comparison of Bovine and Carp Fish Visual Pigment Photo‐Intermediates at Room Temperature

Structural Impact of Selected Retinoids on Model Photoreceptor Membranes

Phototransduction in retinal cones: Analysis of parameter importance

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