cGMP phosphodiesterase dependent light-induced scattering changes in suspensions of retinal disk membranes

Bennett, Nelly; Clerc, Armel

doi:10.1021/bi00121a039

Cited by 5 publications

(8 citation statements)

References 35 publications

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“…This signal we shall term "PDE signal", anticipating its detailed stoichiometric and kinetic analysis. Available data identify it with the fast phase reported in previous phosphodiesterase-dependent turbidity investigations (Bennett & Clerc, 1992; note that turbidity changes are of opposite polarity). When PDE but no transducin is added to membranes, a weak flash does not evoke any scattering change.…”

Section: Kdsupporting

confidence: 67%

“…When purified PDE is added to an otherwise identical sample, the same flash evokes a different light-scattering change. It extends over a longer time scale (Figure lA,a), with complex kinetics (Caretta & Stein, 1986;Bennett & Clerc, 1992). On a short time scale, a fast signal of positive polarity can be clearly separated (Figure lB,a).…”

Section: Kdmentioning

confidence: 94%

“…Titration with G* leads to an increasing level of G*PDE,b until the population of "first" binding sites on PDE is saturated by equimolar G*. Addition of excess transducin evokes the dissociation signal and a slower light-scattering change, which is related to the G*2PDE complex (Caretta & Stein, 1986;Bennett & Clerc, 1992), but slower than its formation ( Figures 1A,a and 5B).…”

Section: The Pde Signal May Monitor the Transition Into The Partiallymentioning

confidence: 99%

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G-protein-effector coupling: A real-time light-scattering assay for transducin-phosphodiesterase interaction

Heck¹,

Hofmann²

1993

Biochemistry

100

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We describe a real-time assay for the interaction of the G-protein of retinal rods, transducin (Gt), with its effector, cGMP phosphodiesterase (PDE). The purified proteins were recombined with isolated rod disk membranes. Reactions were triggered by flash photolysis of the receptor rhodopsin, and monitored by changes of near-infrared light scattering (LS signals). PDE-related LS signals from membrane preparations have been described by others [Caretta, A., & Stein, P. J. (1986) Biochemistry 25, 2335-2341; Bennett, N., & Clerc, A. (1992) Biochemistry 31, 1858-1866]. In the present study, the previous overall light-scattering (turbidity) approach was improved by a specific optical setup and millisecond time resolution. This allowed isolation of a fast "PDE signal" which monitors Gt-PDE interaction on the membrane and in real time. The assignment to the reaction is made by the following observations: (i) preactivation of PDE by purified G alpha GTP gamma S gradually suppresses the signal; (ii) the peak amplitude of the signal is stoichiometric with respect to both Gt and PDE; and (iii) the rise and delay time fit into a kinetic model for a reaction of activated Gt (G*). A comparative investigation, relating the PDE signal with Gt activation and PDE enzymatic activity, led to the following results: (i) the apparent dissociation constant for the interaction of G* with the first binding site on membrane-bound PDE is less than 2.5 nM; (ii) the time interval between formation of G* and its interaction with PDE is less than 5 ms; (iii) membrane-bound PDE, even in its preactivated form, slows the release of G* from the membrane.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Kdsupporting

confidence: 67%

Section: Kdmentioning

confidence: 94%

Section: The Pde Signal May Monitor the Transition Into The Partiallymentioning

confidence: 99%

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G-protein-effector coupling: A real-time light-scattering assay for transducin-phosphodiesterase interaction

Heck¹,

Hofmann²

1993

Biochemistry

100

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“…2), have been demonstrated in endocrine cells, 46,47 brain cortex, 48-51 retina, [52][53][54][55][56][57][58][59][60][61][62] and other neural tissues. 2), have been demonstrated in endocrine cells, 46,47 brain cortex, 48-51 retina, [52][53][54][55][56][57][58][59][60][61][62] and other neural tissues.…”

Section: Introductionmentioning

confidence: 99%

Intrinsic optical signal imaging of retinal physiology: a review

Yao

Wang

2015

J. Biomed. Opt

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Intrinsic optical signal (IOS) imaging promises to be a noninvasive method for high-resolution examination of retinal physiology, which can advance the study and diagnosis of eye diseases. While specialized optical instruments are desirable for functional IOS imaging of retinal physiology, in depth understanding of multiple IOS sources in the complex retinal neural network is essential for optimizing instrument designs. We provide a brief overview of IOS studies and relationships in rod outer segment suspensions, isolated retinas, and intact eyes. Recent developments of line-scan confocal and functional optical coherence tomography (OCT) instruments have allowed in vivo IOS mapping of photoreceptor physiology. Further improvements of the line-scan confocal and functional OCT systems may provide a feasible solution to pursue functional IOS mapping of human photoreceptors. Some interesting IOSs have already been detected in inner retinal layers, but better development of the IOS instruments and software algorithms is required to achieve optimal physiological assessment of inner retinal neurons.

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“…Retinal rod cyclic GMP (cGMP) phosphodiesterase (PDE), a key component in the phototransduction cascade, is located in the outer segment of the photoreceptor cells (Bennett and Clerc, 1992). Phototransduction begins with the absorption of light by rhodopsin.…”

mentioning

confidence: 99%

Generalized progressive retinal atrophy of Sloughi dogs is due to an 8-bp insertion in exon 21 of the PDE6B gene

Dekomien

Runte

Gödde

et al. 2000

Cytogenet Genome Res

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We investigated the gene encoding the β subunit of cGMP phosphodiesterase (PDE6B) as a candidate for generalized progressive retinal atrophy (gPRA), an autosomal recessively transmitted eye disease in dogs. The PDE6B gene was isolated from a genomic library. Single-strand conformation polymorphism analysis revealed eight intronic variations in different subsets of the 14 dog breeds investigated. In addition, we identified an 8-bp insertion after codon 816 in certain Sloughi dogs. Analysis of PRA-affected and obligatory carrier Sloughis showed that this mutation cosegregates with disease status in a large pedigree. All other exchanges identified were not located in functionally relevant parts of the gene (e.g., in the splice signal consensus sites). In most dog breeds (Labrador retriever, Tibetan mastiff, dachshund, Tibetan terrier, miniature poodle, Australian cattle dog, cocker spaniel, collie, Saarloos wolfhound, Chesapeake Bay retriever, and Yorkshire terrier), PDE6B was excluded as a candidate gene for gPRA because heterozygous allele constellations were detected in diseased animals. Therefore, the PDE6B sequence variations did not segregate together with the mutation(s) causing gPRA. Direct and indirect DNA tests concerning gPRA can be offered now for a variety of different dog breeds.

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cGMP phosphodiesterase dependent light-induced scattering changes in suspensions of retinal disk membranes

Cited by 5 publications

References 35 publications

G-protein-effector coupling: A real-time light-scattering assay for transducin-phosphodiesterase interaction

G-protein-effector coupling: A real-time light-scattering assay for transducin-phosphodiesterase interaction

Intrinsic optical signal imaging of retinal physiology: a review

Generalized progressive retinal atrophy of Sloughi dogs is due to an 8-bp insertion in exon 21 of the PDE6B gene

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