Anion sensitivity and spectral tuning of middle- and long-wavelength-sensitive (MWS/LWS) visual pigments

The long-wavelength-sensitive (LWS) opsins form one of four classes of vertebrate cone visual pigment and exhibit peak spectral sensitivities (λ(max)) that generally range from 525 to 560 nm for rhodopsin/vitamin-A(1) photopigments. Unique amongst the opsin classes, many LWS pigments show anion sensitivity through the interaction of chloride ions with a histidine residue at site 197 (H197) to give a long-wavelength spectral shift in peak sensitivity. Although it has been shown that amino acid substitutions at five sites (180, 197, 277, 285 and 308) are useful in predicting the λ(max) values of the LWS pigment class, some species, such as the elephant shark and most marine mammals, express LWS opsins that possess λ(max) values that are not consistent with this 'five-site' rule, indicating that other interactions may be involved. This study has taken advantage of the natural mutation at the chloride-binding site in the mouse LWS pigment. Through the use of a number of mutant pigments generated by site-directed mutagenesis, a new model has been formulated that takes into account the role of charge and steric properties of the side chains of residues at sites 197 and 308 in the function of the chloride-binding site in determining the peak sensitivity of LWS photopigments.     

Visual pigment: G-protein-coupled receptor for light signals

The visual pigment present in photoreceptor cells is a prototypical G-protein-coupled receptor (GPCR) that receives a light signal from the outer environment using a light-absorbing chromophore, 11-cis-retinal. Through cis-trans isomerization of the chromophore, light energy is transduced into chemical free energy, which is in turn utilized for conformational changes in the protein to activate the retinal G-protein. In combination with site-directed mutagenesis, various spectroscopic and biochemical studies identified functional residues responsible for chromophore binding, color regulation, intramolecular signal transduction and G-protein coupling. Extensive studies reveal that these residues are localized into specific domains of visual pigments, suggesting a highly manipulated molecular architecture in visual pigments. In addition to the recent findings on dysfunctional mutations in patients with retinitis pigmentosa or congenital night blindness, the mechanism of intramolecular signal transduction in visual pigments and their evolutionary relationship are discussed. Received 20 July 1998; received after revision 9 September 1998; accepted 23 September 1998     

The opsins of the vertebrate retina: insights from structural,biochemical, and evolutionary studies

The vertebrate retina contains several classes of visual pigments responsible for such diverse functions as image- and nonimage-forming vision, the entrainment of circadian cycles, and the pupilary light response. With vision being vital to the survival of many species, the elucidation of the structural and biochemical properties of visual pigments has been the focus of a large body of research that has led to rapid advances in the field of photoreception. In this review, the current understanding of the structure, function, biochemistry, and evolution of the opsins that make up the photopigments in the vertebrate retina will be reviewed. These include the rod and cone opsins, melanopsin, RGR, peropsin, and VA-opsin. The goal is to highlight important questions that have been answered and to define some of the remaining questions in the field that will provide future directions for research.     

Spektrale Phototaxis von Planktonrotatorien

Summary The spectral sensitivity of 3 species of planktonic Rotatoria (Asplanchna priodonta, Polyarthra remata, Filinia longiseta) were measured by means of the positive phototactic reaction.Asplanchna andPolyarthra both have a broad maximum sensitivity range around 540 nm and a near maximum at 440 nm. In the UV direction (to 360 nm) the spectral sensitivity increases. In contrast,Filinia has only a maximum of 460 nm in the visible spectrum, but behaves similarly to the other species in the UV region. The ecological significance of these results is discussed, as well as their bearing on a possible dermal light sense.     

Behavioral selection in crayfish correlates with movement of a screening pigment in the eye

Light induces two contrasting behavioral responses in crayfish: attraction at low intensities and withdrawal at high intensities. The aim of our experiments has been to study whether screening pigments of the eye influence the selection of attraction or withdrawal responses. During illumination, screening pigments mask photoreceptor cells, reducing the gain of the visual system. Comparison of the time and light-intensity functions of pigment migration and of attraction and withdrawal responses suggest that pigment migration might influence the selection as well as the latency of the response.     

The structure of scytonemin,an ultraviolet sunscreen pigment from the sheaths of cyanobacteria

Despite knowledge of the existence of the pigment called scytonemin for over 100 years, its structure has remained unsolved until now. This pigment, the first shown to be an effective, photo-stable ultraviolet shield in prokaryotes, is a novel dimeric molecule (molec. wt. 544) of indolic and phenolic subunits and is known only from the sheaths enclosing the cells of cyanobacteria. It is probable that scytonemin is formed from a condensation of tryptophan-and phenylpropanoid-derived subunits. The linkage between these units is unique among natural products and this novel ring structure is here termed the scytoneman skeleton. Scytonemin absorbs strongly and broadly in the spectral region 325–425 nm (UV-A-violet-blue, with an in vivo maximum at 370 nm). However, there is also major absorption in the UV-C ( _max=250nm) and UV-B (280–320 nm). The pigment has been recently shown to provide significant protection to cyanobacteria against damage by ultraviolet radiation. The pigment occurs in all phylogenetic lines of sheathed cyanobacteria and possibly represents a UV screening strategy far more ancient than that of plant flavonoids and animal melanins. How diverse organisms deal with UV radiation is considered of vital importance to global ecology.     

On partial and differential bleaching experiments with the visual pigments in a fresh water euryhaline teleost (Etroplus suratensis)

Summary Visual pigment was extracted from a fresh water toleost,Etroplus suratensis and the optical density measured over a range of 300–650 nm. The absorption spectrum indicates the _max at 550 nm and a small hump at 530 nm. Through partial bleaching at 630 nm, it was confirmed that the fish possesses a mixture of 2 visual pigments: the one with the _max at 550 nm predominating over the other.     

A survey of 3-dehydroretinal as a visual pigment chromophore in various species of crayfish and other freshwater crustaceans

Summary 3-Dehydroretinal (vitamin A₂ aldehyde) was found in the eyes of three species among 10 species of freshwater crayfish examined. Since dark-adapted eyes contained the 11-cis form of 3-dehydroretinal, this compound must be the chromophore of the visual pigment. 3-Dehydroretinal always coexisted with retinal (vitamin A₁ aldehyde), indicating the presence of a rhodopsin-porphyropsin visual pigment system.     

Alternariol, a dibenzopyrone mycotoxin of Alternaria spp., is a new photosensitizing and DNA cross-linking agent

The mycotoxin alternariol (3,4',5-trihydroxy-6'-methyldibenzo [a] pyrone) but not alternariol monomethyl ether (3,4'-dihydroxy-5-methoxy-6'-methyldibenzo [a] pyrone) is phototoxic to Escherichia coli in the presence of near UV light (320-400 nm). The phototoxicity bioassays with a DNA repair-deficient mutant of E. coli suggested that DNA may be the molecular target for photo-induced toxicity of alternariol. Interactions between alternariol and double-stranded, supercoiled DNA suggest that alternariol interacts with DNA by intercalation. No DNA breakage was detected in this system; however, alternariol forms a complex and cross-links double-stranded DNA in near UV light. These results suggest that alternariol is a new phototoxic, DNA-intercalating agent and is a DNA cross-linking mycotoxin in near UV light.     

Experimental studies of schistosomal pigment from Schistosoma japonicum.

The schistosomal and malarial pigments were distinguishable before and after extraction from the host liver. Presence of iron in both pigments was ascertained by the elemental X-ray analysis. Histochemically, however, schistosomal pigment was similar to that of malarial pigment.     

Experimental studies of schistosomal pigment fromSchistosoma japonicum

Summary The schistosomal and malarial pigments were distinguishable before and after extraction from the host liver. Presence of iron in both pigments was ascertained by the elemental X-ray analysis. Histochemically, however, schistosomal pigment was similar to that of malarial pigment.     

Pigment formation in sensory cells ofAplysia l

Summary The yellow-brown pigment present in the sensory cells ofAplysia limacina was studied using light and electron microscopy. The ultrastructure, the high carotenoid content and the presence in neurons for which a turnover process has been hypothesized, indicate that these pigments are cytosomes, organelles involved in the production of energy in anaerobiosis.This investigation was conducted at the Zoological Station of Naples.     

Mitochondrial quality control in AMD: does mitophagy play a pivotal role?

Age-related macular degeneration (AMD) is the predominant cause of visual loss in old people in the developed world, whose incidence is increasing. This disease is caused by the decrease in macular function, due to the degeneration of retinal pigment epithelium (RPE) cells. The aged retina is characterised by increased levels of reactive oxygen species (ROS), impaired autophagy, and DNA damage that are linked to AMD pathogenesis. Mitophagy, a mitochondria-specific type of autophagy, is an essential part of mitochondrial quality control, the collective mechanism responsible for this organelle’s homeostasis. The abundance of ROS, DNA damage, and the excessive energy consumption in the ageing retina all contribute to the degeneration of RPE cells and their mitochondria. We discuss the role of mitophagy in the cell and argue that its impairment may play a role in AMD pathogenesis. Thus, mitophagy as a potential therapeutic target in AMD and other degenerative diseases is as well explored.     

A visual pigment of the sturgeon retina

The visual pigments of hybrid sturgeon (a cross between Acipenser ruthenus (male) and Huso huso (female) have been studied both by the methods of incomplete partial bleaching and HPLC analysis. On the basis of the results obtained the relationship between the structure of opsins and the spectral characteristics of visual pigments is discussed.     

Photoreactivation of UV damage in cultured Drosophila cells

Cell survival and photoreactivation of 254 nm ultraviolet (UV) light damage in a wild type Drosophila cell line was assayed by colony formation in liquid medium. Fo, Fq, and extrapolation number for the exponential portion of survival curves are 21 J/m2, 3.6 J/m2, and 1.5 for non-photoreactivated cells and 110 J/m2, 11.2 J/m2, and 1.3 for those exposed to photoreactivating light. Maximal photoreactivation occurs at the 100 J/m2 region of the curve. At 10 and 50% survival, 75-80% of the UV damage was photoreactivable.     

UV guided dendritic growth patterns and the networking of melanocytes

Whole skin organ cultures of vitiliginous, skin show that the marginal melanocytes are highly sensitive to a pulse of UV exposure (210–380 nm) during the G₂ phase of the cell cycle, as seen by prominent dendricity. Melanocytes are highly dendritic in the epidermis overlying rapidly growing tumors, as well as within proliferative lesions such as basal cell carcinomas and aggressive seborrheic keratosis. In the organ cultures the dendrites extend towards the source of UV, i.e. the surface, while the main body lies along the basement membrane. The epidermal melanocytes overlying tumors lie, almost vertically, dendrites aligned towards the underlying tumor on one side and the surface on the other. Within tumors dendritic elongation is guided by mitotic and PCNA positive (S-phase) tumor cells, which are a source of ultraweak UV emissions in the range of 210–330 nm. These observations indicate that ultraweak biophoton emissions from neighbouring cells can simulate environmental cues and contribute to the plasticity of networks such as the melanocytes or the visual pathways.     

Insights into acylphosphatase structure and catalytic mechanism

Acylphosphatase is one of the smallest enzymes known (about 98 amino acid residues). It is present in organs and tissues of vertebrate species as two isoenzymes sharing over 55% of sequence homology; these appear highly conserved in differing species. The two isoenzymes can be involved in a number of physiological processes, though their effective biological function is not still certain. The solution and crystal structures of different isoenzymes are known, revealing a close packed protein with a fold similar to that shown by other phosphate-bind ing proteins. The structural data, together with an extended site-directed mutagenesis investigation, led to the identification of the residues involved in enzyme catalysis. However, it appears unlikely that these residues are able to perform the full catalytic cycle: a substrate-assisted catalytic mechanism has therefore been proposed, in which the phosphate moiety of the substrate could act as a nucleophile activating the catalytic water molecule. Received 12 November 1996; accepted 27 November 1996     

Studies on the reconstitution ofP. clarkii blue carotenoprotein from its isolated subunits

Summary A blue carotenoid-protein complex (_max 635 nm) was extracted and purified from the carapace of the crayfishProcambarus clarkii. The complex was further liberated from astaxanthin, its prostetic group, causing dissociation into apoprotein subunits. Reconstitution of the complex from the various sub-units (isolated by chromatofocusing) plus astaxanthin was attempted. Apoprotein-size pigments of rose-purple color (_max 545 nm) were obtained. It was found that both monomers are required in order to a blue complex fairly similar in structure ot the native one. However, the native conformation was not completely recovered, as indicated by some differences in the UV spectrum.     

Photolyse ultraviolette et photoréactivation des bactéries — Aspects de dépolymérisation et repolymérisation des acides nucléiques bactériens

Summary Electrophoretic analysis of bacterial substance proves the departure, from UV irradiated microbes screened from light for a certain time, of substances which were responsible for increasing the charge density by surface unit during the first moments which follow the irradiation by UV rays. Spectrophotometric analysis of liquids having contained these bacilli proves concommitantly the diffusion of those substances from the bacterial substance to the aqueous phase and reveals their nature. The photoreactivity which is possible during the first moments following the UV irradiation of bacilli, due to the presence of the fragments of depolymerisation of nucleic acid macromolecule in the bacterial substance, becomes impossible after the departure (diffusion) of those fragments to the aqueous phase. These observations seem to show that the phenomenon of photo-reactivity is connected with the repolymerisation of the macromolecule primitively divided of desoxyribonucleic acid.     

2-Methoxybenzoyl phosphate: a new substrate for continuous fluorimetric and spectrophotometric acyl phosphatase assays

A new aromatic acyl phosphate, 2-methoxybenzoyl phosphate, has been synthesized. The compound shows an intrinsic fluorescence; it displays an intense emission band at 390 nm upon excitation in the near UV region. This band practically disappears after hydrolysis of the product. On the other hand, the product displays differences in the near UV absorption spectra measured before and after hydrolysis. The at 301 nm is 2720 M^–1 cm^–1, a value that is 4.3-fold higher than that of benzoyl phosphate (the usual substrate for acylphosphatase assay) at 283 nm. The main kinetic parameters of three different acylphosphatase molecular forms (the muscular isoenzyme and two subtypes of the organ common isoenzyme) were determined using both benzoyl phosphate and 2-methoxybenzoyl phosphate as substrates, and then compared. These kinetic data and the UV absorption and fluorescence properties of 2-methoxybenzoyl phosphate sugest that this compound has better substrate features than benzoyl phosphate, and can be used for both high sensitivity continuous fluorimetric and UV absorption spectrophotometric assays of acylphosphatase.