In vitro assay for trans-phosphorylation of rhodopsin by rhodopsin kinase

Biochemistry
J RimDaniel D Oprian

Abstract

Trans-phosphorylation of rhodopsin refers to a reaction in which a rhodopsin kinase molecule that has been activated by a light-activated rhodopsin molecule collides with and phosphorylates a second molecule of rhodopsin that has not been activated by light. It has been invoked as a mechanism for high-gain phosphorylation, a phenomenon that is observed at low bleaching levels where up to several hundred moles of phosphate are added to the rhodopsin pool per mole of photolyzed rhodopsin. Trans-phosphorylation is an appealing mechanism to propose for high-gain phosphorylation, but it has not been tested directly because of the difficulty inherent in unambiguous identification of light-activated and dark forms of rhodopsin present in the same reaction mixture. We report here a direct assay for trans-phosphorylation of rhodopsin. The assay is based on the use of a split receptor mutant of rhodopsin, SR(1-4/5-7), in which the fully functional protein is assembled from two separately expressed fragments. Because of different electrophoretic mobilities, SR(1-4/5-7) and wild-type rhodopsin can be monitored independently for phosphorylation while in the same reaction mixture. Thus, if wild-type rhodopsin is exposed to light and then inc...Continue Reading

References

Mar 1, 1992·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·P A Hargrave, J H McDowell
Aug 16, 1996·The Journal of Biological Chemistry·B M BinderV Y Arshavsky
Jan 15, 1972·FEBS Letters·H Kühn, W J. Dreyer
Nov 1, 1963·The Journal of General Physiology·R G MATTHEWSG WALD
Sep 26, 1988·FEBS Letters·Charles FowlesM Akhtar

❮ Previous
Next ❯

Citations

May 14, 2003·Progress in Retinal and Eye Research·Tadao MaedaKrzysztof Palczewski
Apr 5, 2002·The Biochemical Journal·Nina E M McCarthy, Muhammad Akhtar
Oct 6, 1997·The Journal of General Physiology·W XiongK Yau
Jul 17, 1998·Vision Research·J B HurleyG A Niemi
Oct 13, 2005·The Journal of Biological Chemistry·Guang W ShiJeannie Chen

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.