Transducin (G
t) is a
protein
Proteins are large biomolecules and macromolecules that comprise one or more long chains of amino acid residues. Proteins perform a vast array of functions within organisms, including catalysing metabolic reactions, DNA replication, respo ...
naturally
expressed in
vertebrate
Vertebrates () comprise all animal taxa within the subphylum Vertebrata () ( chordates with backbones), including all mammals, birds, reptiles, amphibians, and fish. Vertebrates represent the overwhelming majority of the phylum Chordata, ...
retina
The retina (from la, rete "net") is the innermost, light-sensitive layer of tissue of the eye of most vertebrates and some molluscs. The optics of the eye create a focused two-dimensional image of the visual world on the retina, which then ...
rods and cones and it is very important in
vertebrate phototransduction. It is a type of
heterotrimeric G-protein with different α subunits in rod and cone photoreceptors.
Light leads to conformational changes in
rhodopsin
Rhodopsin, also known as visual purple, is a protein encoded by the RHO gene and a G-protein-coupled receptor (GPCR). It is the opsin of the rod cells in the retina and a light-sensitive receptor protein that triggers visual phototransduction ...
, which in turn leads to the activation of transducin. Transducin activates
phosphodiesterase
A phosphodiesterase (PDE) is an enzyme that breaks a phosphodiester bond. Usually, ''phosphodiesterase'' refers to cyclic nucleotide phosphodiesterases, which have great clinical significance and are described below. However, there are many oth ...
, which results in the breakdown of
cyclic guanosine monophosphate
Cyclic guanosine monophosphate (cGMP) is a cyclic nucleotide derived from guanosine triphosphate (GTP). cGMP acts as a second messenger much like cyclic AMP. Its most likely mechanism of action is activation of intracellular protein kinases in re ...
(cGMP). The intensity of the flash response is directly proportional to the number of transducin activated.
Function in phototransduction
Transducin is activated by
metarhodopsin II, a conformational change in
rhodopsin
Rhodopsin, also known as visual purple, is a protein encoded by the RHO gene and a G-protein-coupled receptor (GPCR). It is the opsin of the rod cells in the retina and a light-sensitive receptor protein that triggers visual phototransduction ...
caused by the
absorption
Absorption may refer to:
Chemistry and biology
* Absorption (biology), digestion
**Absorption (small intestine)
*Absorption (chemistry), diffusion of particles of gas or liquid into liquid or solid materials
*Absorption (skin), a route by which ...
of a
photon
A photon () is an elementary particle that is a quantum of the electromagnetic field, including electromagnetic radiation such as light and radio waves, and the force carrier for the electromagnetic force. Photons are massless, so they always ...
by the rhodopsin moiety
retinal
Retinal (also known as retinaldehyde) is a polyene chromophore. Retinal, bound to proteins called opsins, is the chemical basis of visual phototransduction, the light-detection stage of visual perception (vision).
Some microorganisms use retin ...
.
The light causes isomerization of retinal from 11-cis to all-trans. Isomerization causes a change in the
opsin
Animal opsins are G-protein-coupled receptors and a group of proteins made light-sensitive via a chromophore, typically retinal. When bound to retinal, opsins become Retinylidene proteins, but are usually still called opsins regardless. Most pro ...
to become metarhodopsin II. When metarhodopsin activates transducin, the
guanosine diphosphate
Guanosine diphosphate, abbreviated GDP, is a nucleoside diphosphate. It is an ester of pyrophosphoric acid with the nucleoside guanosine. GDP consists of a pyrophosphate group, a pentose sugar ribose, and the nucleobase guanine.
GDP is the product ...
(GDP) bound to the α subunit (T
α) is exchanged for
guanosine triphosphate
Guanosine-5'-triphosphate (GTP) is a purine nucleoside triphosphate. It is one of the building blocks needed for the synthesis of RNA during the transcription process. Its structure is similar to that of the guanosine nucleoside, the only diffe ...
(GTP) from the cytoplasm. The α subunit dissociates from the βγ subunits (T
βγ). Activated transducin α-subunit activates cGMP phosphodiesterase. cGMP
phosphodiesterase
A phosphodiesterase (PDE) is an enzyme that breaks a phosphodiester bond. Usually, ''phosphodiesterase'' refers to cyclic nucleotide phosphodiesterases, which have great clinical significance and are described below. However, there are many oth ...
breaks down cGMP, an intracellular
second messenger
Second messengers are intracellular signaling molecules released by the cell in response to exposure to extracellular signaling molecules—the first messengers. (Intercellular signals, a non-local form or cell signaling, encompassing both first me ...
which opens cGMP-gated cation channels. Phosphodiesterase hydrolyzes cGMP to 5’-GMP. Decrease in cGMP concentration leads to decreased opening of cation channels and subsequently, hyperpolarization of the
membrane potential
Membrane potential (also transmembrane potential or membrane voltage) is the difference in electric potential between the interior and the exterior of a biological cell. That is, there is a difference in the energy required for electric charges ...
.
Transducin is deactivated when the α-subunit-bound GTP is hydrolyzed to GDP. This process is accelerated by a complex containing an RGS (''Regulator of G-protein signaling'')-protein and the gamma-subunit of the effector, cyclic GMP Phosphodiesterase.
Mechanism of activation
The T
α subunit of transducin contains three functional domains: one for rhodopsin/T
βγ interaction, one for GTP binding, and the last for activation of cGMP phosphodiesterase.
Although the focus for phototransduction is on T
α, T
βγ is crucial for rhodopsin to bind to transducin.
The rhodopsin/T
βγ binding domain contains the
amino
In chemistry, amines (, ) are compounds and functional groups that contain a basic nitrogen atom with a lone pair. Amines are formally derivatives of ammonia (), wherein one or more hydrogen atoms have been replaced by a substituent ...
and
carboxyl terminal of the T
α. The amino terminal is the site of interaction for rhodopsin while the carboxyl terminal is that for T
βγ binding. The amino terminal might be anchored or in close proximity to the carboxyl terminal for activation of the transducin molecule by rhodopsin.
Interaction with photolyzed rhodopsin opens up the GTP-binding site to allow for rapid exchange of GDP for GTP. The binding site is in the closed conformation in the absence of photolyzed rhodopsin. Normally in the closed conformation, an
α-helix
The alpha helix (α-helix) is a common motif in the secondary structure of proteins and is a right hand-helix conformation in which every backbone N−H group hydrogen bonds to the backbone C=O group of the amino acid located four residues e ...
located near the binding site is in a position which hinders the GTP/GDP exchange. A conformational change of the T
α by photolyzed rhodopsin causes the tilting of the helix, opening the GTP-binding site.
Once GTP has been exchanged for GDP, the GTP-T
α complex undergoes two major changes: dissociation from photolyzed rhodopsin and the T
βγ subunit and exposure of the phosphodiesterase (PDE) binding site for interaction with latent PDE. The conformational changes initiated in the transducin by binding of GTP are transmitted to the PDE binding site and cause it to be exposed for binding to PDE. The GTP-induced conformational changes could also disrupt the rhodopsin/T
βγ binding site and lead to dissociation from the GTP-T
α complex.
The Tβγ complex
An underlying assumption for G-proteins is that α, β, and γ subunits are present in the same concentration. However, there is evidence that there are more T
β and T
γ than T
α in rod outer segments (ROS).
The excess T
β and T
γ have been concluded to be floating freely around in the ROS, though it cannot be associated with the T
α at any given time. One possible explanation for the excess T
βγ is increased availability for T
α to rebind. Since T
βγ is crucial for the binding of transducin, reacquisition of the heterotrimeric conformation could lead to more rapid binding to another GTP molecule and thus faster phototransduction.
Though T
βγ has been mentioned to be crucial for T
α binding to rhodopsin, there is also evidence that T
βγ may have a crucial, possibly direct role in nucleotide exchange than previously thought. Rhodopsin was found to specifically cause a conformational switch in the carboxyl terminal of the T
γ subunit. This change ultimately regulates the allosteric nucleotide exchange on the T
α. This domain could serve as a major area for interactions with rhodopsin and for rhodopsin to regulate nucleotide exchange on the T
α. Activation of the G protein transducin by rhodopsin was thought to proceed by the lever mechanism. Rhodopsin-binding causes helix formation at the carboxyl terminal on the T
γ and brings the T
γ carboxyl and T
α. Carboxyl terminals closer together to facilitate nucleotide exchange.
Mutations in this domain abolish rhodopsin-transducin interaction. This conformational switch in the T
γ may be preserved in the G protein γ subunit family.
Interaction with cGMP phosphodiesterase and deactivation
Transducin activation ultimately results in stimulation of the biological effector molecule cGMP phosphodiesterase, an oligomer with α, β and two inhibitory γ subunits. The α and β subunits are the larger molecular weight subunits and make up the catalytic moiety of PDE.
In the phototransduction system, GTP-bound-T
α binds to the γ subunit of PDE. There are two proposed mechanisms for the activation of PDE. The first proposes that the GTP-bound-T
α releases the PDE γ subunit from the catalytic subunits in order to activate hydrolysis.
The second more likely mechanism proposes that binding causes a positional shift of the γ subunit, allowing better accessibility of the catalytic subunit for cGMP hydrolysis. The GTPase activity of T
α hydrolyzes GTP to GDP and changes the conformation of the T
α subunit, increasing its affinity to bind to the α and β subunits on the PDE. The binding of T
α to these larger subunits results in another conformational change in PDE and inhibits the hydrolysis ability of the catalytic subunit. This binding site on the larger molecular subunit may be immediately adjacent to the T
α binding site on the γ subunit.
Although the traditional mechanism involves activation of PDE by GTP-bound T
α, GDP-bound T
α has also been demonstrated to have the ability to activate PDE. Experiments of PDE activation in the dark (without the presence of GTP) show small but reproducible PDE activation.
This can be explained by the activation of PDE by free GDP-bound T
α. PDE γ subunit affinity for GDP-bound T
α, however, seems to be about 100-fold smaller than for GTP-bound T
α.
The mechanism by which GDP-bound T
α activates PDE remains unknown however, it is speculated to be similar to the activation of PDE by GTP-bound T
α.
In order to prevent activation of PDE in the dark, the concentration of GDP-bound T
α should be kept to a minimum. This job seems to fall to the T
βγ to keep the GDP-bound T
α bound in the form of holotransducin.
For deactivation, hydrolysis of the bound GTP by the T
α is necessary for T
α deactivation and returning the transducin to its basal from. However, simple hydrolysis of GTP may not necessarily be enough to deactivate PDE. T
βγ comes into play here again with an important role in PDE deactivation.
The addition of T
βγ facilitates inhibition of the PDE catalytic moiety because it binds with the T
α-GTP complex. The reassociated form of transducin is not able to bind to PDE any longer. This frees PDE to recouple to photolyzed rhodopsin and return PDE to its initial state to await activation by another GTP bound T
α.
Genes
* ,,
References
External links
*
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G proteins