View source for Transducin

{{short description|Heterotrimeric G protein involved in vertebrate phototransduction}}
[[Image:Rhodopsin-transducin.png|thumb|right|[[Sensory rhodopsin II]] (rainbow colored) embedded in a [[lipid bilayer]] (heads red and tails blue) with Transducin below it. G<sub>t</sub>α is colored red, G<sub>t</sub>β blue, and G<sub>t</sub>γ yellow. There is a bound [[Guanosine diphosphate|GDP]] molecule in the G<sub>t</sub>α-subunit and a bound retinal (black) in the rhodopsin. The [[amino-terminus|N-terminus]] of rhodopsin is red and the [[C-terminus]] blue. Anchoring of transducin to the membrane has been drawn in black.]]

'''Transducin''' (G<sub>t</sub>) is a [[protein]] naturally [[gene expression|expressed]] in [[vertebrate]] [[retina]] rods and cones and it is very important in [[visual phototransduction|vertebrate phototransduction]]. It is a type of [[heterotrimeric G-protein]] with different α subunits in rod and cone photoreceptors.<ref name="pmid3529395">{{cite journal |vauthors=Lerea CL, Somers DE, Hurley JB, Klock IB, Bunt-Milam AH |title=Identification of specific transducin alpha subunits in retinal rod and cone photoreceptors |journal=Science |volume=234 |issue=4772 |pages=77–80 |date=October 1986 |pmid=3529395 |doi= 10.1126/science.3529395}}</ref>

Light leads to conformational changes in the G protein–coupled receptor [[rhodopsin]], which in turn leads to the activation of transducin. Transducin activates [[phosphodiesterase]], which results in the breakdown of [[cyclic guanosine monophosphate]] (cGMP). The intensity of the flash response is directly proportional to the number of transducin activated.

==Function in phototransduction==
Transducin is activated by [[rhodopsin|metarhodopsin II]], a conformational change in [[rhodopsin]] caused by the [[absorption (electromagnetic radiation)|absorption]] of a [[photon]] by the rhodopsin moiety [[retinal]].<ref name="pmid8466475">{{cite journal |vauthors=Hargrave PA, Hamm HE, Hofmann KP |title=Interaction of rhodopsin with the G-protein, transducin |journal=BioEssays |volume=15 |issue=1 |pages=43–50 |date=January 1993 |pmid=8466475 |doi=10.1002/bies.950150107 |s2cid=9487394 }}</ref><ref name="pmid16989885">{{cite journal |vauthors=Downs MA, Arimoto R, Marshall GR, Kisselev OG |title=G-protein alpha and beta-gamma subunits interact with conformationally distinct signaling states of rhodopsin |journal=Vision Res. |volume=46 |issue=27 |pages=4442–8 |date=December 2006 |pmid=16989885 |doi=10.1016/j.visres.2006.07.021 |doi-access=free }}</ref> The light causes isomerization of retinal from 11-''cis'' to all-''trans''. Isomerization causes a change in the [[opsin]] to become metarhodopsin II. When metarhodopsin activates transducin, the [[guanosine diphosphate]] (GDP) bound to the [[G alpha subunit|α subunit]] (T<sub>α</sub>) is exchanged for [[guanosine triphosphate]] (GTP) from the cytoplasm. The α subunit dissociates from the [[G_beta-gamma_complex|βγ subunit]]s (T<sub>βγ</sub>). Activated transducin α-subunit activates [[Cyclic guanosine monophosphate|cGMP]] [[phosphodiesterase]].<ref>{{cite journal | last1 = Fung | first1 = BKK | last2 = Hurley | first2 = JB | last3 = Stryer | first3 = L | year = 1981 | title = Flow of information in the light-triggered cyclic nucleotide cascade of vision | journal = Proceedings of the National Academy of Sciences of the United States of America | volume = 78 | issue = 1| pages = 152–156 | pmid = 6264430 | pmc=319009 | doi=10.1073/pnas.78.1.152| doi-access = free }}</ref> cGMP phosphodiesterase breaks down cGMP, an intracellular [[second messenger]] which opens cGMP-gated cation channels. Phosphodiesterase hydrolyzes cGMP to [[Guanosine monophosphate|5’-GMP]]. Decrease in cGMP concentration leads to decreased opening of cation channels and subsequently hyperpolarization of the [[membrane potential]].

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<sub>α</sub> subunit of transducin contains three functional domains: one for rhodopsin/T<sub>βγ</sub> interaction, one for GTP binding, and the last for activation of cGMP phosphodiesterase.

There are different isoforms of T<sub>α,</sub> seen in rod and cone cells. However, the isoforms exhibit functional interchangeability in the phototransduction cascade and shouldn't solely account for differences in light sensitivity. <ref>{{Cite journal |last=Deng |first=Wen-Tao |date=October 20, 2009 |title=Functional Interchangeability of Rod and Cone Transducin α-Subunits |journal=Proceedings of the National Academy of Sciences of the United States of America |volume=106 |pages=2}}</ref> Although the focus for phototransduction is on T<sub>α</sub>, T<sub>βγ</sub> is crucial for rhodopsin to bind to transducin.<ref>{{Cite journal
 | last1 = Fung | first1 = B. K.
 | title = Characterization of transducin from bovine retinal rod outer segments. I. Separation and reconstitution of the subunits
 | journal = The Journal of Biological Chemistry
 | volume = 258
 | issue = 17
 | pages = 10495–10502
 | year = 1983
 | doi = 10.1016/S0021-9258(17)44483-8
 | pmid = 6136509
| doi-access = free
 }}</ref><ref name="pmid12679015">{{Cite journal
 | last1 = Kisselev | first1 = O. G.
 | last2 = Downs | first2 = M. A.
 | title = Rhodopsin controls a conformational switch on the transducin gamma subunit
 | journal = Structure
 | volume = 11
 | issue = 4
 | pages = 367–373
 | year = 2003
 | pmid = 12679015
 | doi=10.1016/s0969-2126(03)00045-5
| doi-access = free
 }}</ref> The rhodopsin/T<sub>βγ</sub> binding domain contains the [[N-terminus|amino]] and [[C-terminus|carboxyl terminal]] of the T<sub>α</sub>. The amino terminal is the site of interaction for rhodopsin while the carboxyl terminal is that for T<sub>βγ</sub> binding. The amino terminal might be anchored or in close proximity to the carboxyl terminal for activation of the transducin molecule by rhodopsin.<ref name="pmid3038611">{{Cite journal
 | doi = 10.1016/0014-5793(87)80867-0
 | last1 = Hingorani | first1 = V. N.
 | last2 = Ho | first2 = Y. K.
 | title = A structural model for the alpha-subunit of transducin. Implications of its role as a molecular switch in the visual signal transduction mechanism
 | journal = FEBS Letters
 | volume = 220
 | issue = 1
 | pages = 15–22
 | year = 1987
 | pmid = 3038611
| s2cid = 33255299 | doi-access = free
 }}</ref>

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 [[alpha helix|α-helix]] located near the binding site is in a position which hinders the GTP/GDP exchange. A conformational change of the T<sub>α</sub> by photolyzed rhodopsin causes the tilting of the helix, opening the GTP-binding site.

Once GTP has been exchanged for GDP, the GTP-T<sub>α</sub> complex undergoes two major changes: dissociation from photolyzed rhodopsin and the T<sub>βγ</sub> 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<sub>βγ</sub> binding site and lead to dissociation from the GTP-T<sub>α</sub> complex.<ref name="pmid3038611"/>

===The T<sub>βγ</sub> 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<sub>β</sub> and T<sub>γ</sub> than T<sub>α</sub> in rod outer segments (ROS).<ref name="pmid16895762">{{Cite journal
 | last1 = Clack | first1 = J. W.
 | last2 = Springmeyer | first2 = M. L.
 | last3 = Clark | first3 = C. R.
 | last4 = Witzmann | first4 = F. A.
 | title = Transducin subunit stoichiometry and cellular distribution in rod outer segments
 | doi = 10.1016/j.cellbi.2006.06.007
 | journal = Cell Biology International
 | volume = 30
 | issue = 10
 | pages = 829–835
 | year = 2006
 | pmid = 16895762
| s2cid = 20446982
 }}</ref> The excess T<sub>β</sub> and T<sub>γ</sub> have been concluded to be floating freely around in the ROS, though it cannot be associated with the T<sub>α</sub> at any given time. One possible explanation for the excess T<sub>βγ</sub> is increased availability for T<sub>α</sub> to rebind. Since T<sub>βγ</sub> 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.<ref name="pmid16895762"/>

Though T<sub>βγ</sub> has been mentioned to be crucial for T<sub>α</sub> binding to rhodopsin, there is also evidence that T<sub>βγ</sub> 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<sub>γ</sub> subunit. This change ultimately regulates the allosteric nucleotide exchange on the T<sub>α</sub>. This domain could serve as a major area for interactions with rhodopsin and for rhodopsin to regulate nucleotide exchange on the T<sub>α</sub>. Activation of the G protein transducin by rhodopsin was thought to proceed by the lever mechanism.<ref>{{Cite journal
 | last1 = Bourne | first1 = H. R.
 | last2 = Iiri | first2 = T.
 | last3 = Farfel | first3 = Z.
 | title = G-protein diseases furnish a model for the turn-on switch
 | journal = Nature
 | volume = 394
 | issue = 6688
 | pages = 35–38
 | doi = 10.1038/27831
 | year = 1998
 | pmid = 9665125
| s2cid = 204998245
 }}</ref><ref>{{Cite journal
 | last1 = Rondard | first1 = P.
 | last2 = Iiri | first2 = T.
 | last3 = Srinivasan | first3 = S.
 | last4 = Meng | first4 = E.
 | last5 = Fujita | first5 = T.
 | last6 = Bourne | first6 = H. R.
 | title = Mutant G protein α subunit activated by Gβγ: A model for receptor activation?
 | doi = 10.1073/pnas.101136198
 | journal = Proceedings of the National Academy of Sciences
 | volume = 98
 | issue = 11
 | pages = 6150–6155
 | pmc = 33437
 | year = 2001
 | pmid = 11344266
| doi-access = free
 }}</ref> Rhodopsin-binding causes helix formation at the carboxyl terminal on the T<sub>γ</sub> and brings the T<sub>γ</sub> carboxyl and T<sub>α</sub>. Carboxyl terminals closer together to facilitate nucleotide exchange. T<sub>α</sub> can accelerate the rate of activation of light-off induced [[Protein kinase A|Protein Kinase A]] due to binding to [[rhodopsin]].<ref>{{Cite journal |last=Sato |first=Shinya |date=October 27, 2020 |title=Rhodopsin-mediated light-off-induced protein kinase A activation in mouse rod photoreceptor cells |url=https://www.jstor.org/stable/26970649 |journal=Proceedings of the National Academy of Sciences of the United States of America |volume=117 |issue=43 |pages=1}}</ref> As well as, transducin achieves full functional activation upon binding to activated [[rhodopsin]].<ref>{{Cite journal |last=Ernst |first=Oliver |date=June 26, 2006 |title=Monomeric G Protein-Coupled Receptor Rhodopsin in Solution Activates Its G Protein Transducin at the Diffusion Limit |url=https://www.jstor.org/stable/25436029?searchText=transducin&searchUri=%2Faction%2FdoBasicSearch%3FQuery%3Dtransducin%26so%3Drel&ab_segments=0%2Fbasic_search_gsv2%2Fcontrol&refreqid=fastly-default%3Ad470b8e3965af235615409f57b3403da |journal=Proceedings of the National Academy of Sciences of the United States of America |volume=104 |issue=26 |pages=5}}</ref>

Mutations in this domain abolish rhodopsin-transducin interaction. This conformational switch in the T<sub>γ</sub> may be preserved in the G protein γ subunit family.<ref name="pmid12679015"/>

===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.<ref>{{Cite journal 
| last1 = Deterre | first1 = P. 
| last2 = Bigay | first2 = J. 
| last3 = Forquet | first3 = F. 
| last4 = Robert | first4 = M. 
| last5 = Chabre | first5 = M. 
| title = CGMP phosphodiesterase of retinal rods is regulated by two inhibitory subunits 
| journal = Proceedings of the National Academy of Sciences of the United States of America 
| volume = 85 
| issue = 8 
| pages = 2424–2428 
| year = 1988 
| pmid = 2833739 
| pmc = 280009 | doi=10.1073/pnas.85.8.2424
| doi-access = free 
}}</ref> 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<sub>α</sub> 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<sub>α</sub> releases the PDE γ subunit from the catalytic subunits in order to activate hydrolysis.<ref name="pmid2538446">{{Cite journal 
| last1 = Kroll | first1 = S. 
| last2 = Phillips | first2 = W. J. 
| last3 = Cerione | first3 = R. A. 
| title = The regulation of the cyclic GMP phosphodiesterase by the GDP-bound form of the alpha subunit of transducin 
| journal = The Journal of Biological Chemistry 
| volume = 264 
| issue = 8 
| pages = 4490–4497 
| year = 1989 
| doi = 10.1016/S0021-9258(18)83770-X 
| pmid = 2538446
| doi-access = free 
}}</ref> 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<sub>α</sub> hydrolyzes GTP to GDP and changes the conformation of the T<sub>α</sub> subunit, increasing its affinity to bind to the α and β subunits on the PDE. The binding of T<sub>α</sub> 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<sub>α</sub> binding site on the γ subunit.<ref name="pmid2538446"/>

Although the traditional mechanism involves activation of PDE by GTP-bound T<sub>α</sub>, GDP-bound T<sub>α</sub> 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.<ref name="pmid8143750">{{Cite journal
 | last1 = Kutuzov | first1 = M.
 | last2 = Pfister | first2 = C.
 | title = Activation of the retinal cGMP-specific phosphodiesterase by the GDP-loaded alpha-subunit of transducin
 | journal = European Journal of Biochemistry
 | volume = 220
 | issue = 3
 | pages = 963–971
 | year = 1994
 | pmid = 8143750
 | doi=10.1111/j.1432-1033.1994.tb18700.x
| doi-access = free
 }}</ref> This can be explained by the activation of PDE by free GDP-bound T<sub>α</sub>. PDE γ subunit affinity for GDP-bound T<sub>α</sub>, however, seems to be about 100-fold smaller than for GTP-bound T<sub>α</sub>.<ref>{{Cite journal
 | last1 = Bennett | first1 = N.
 | last2 = Clerc | first2 = A.
 | title = Activation of cGMP phosphodiesterase in retinal rods: Mechanism of interaction with the GTP-binding protein (transducin)
 | journal = Biochemistry
 | volume = 28
 | issue = 18
 | pages = 7418–7424
 | year = 1989
 | pmid = 2554970
 | doi=10.1021/bi00444a040
}}</ref> The mechanism by which GDP-bound T<sub>α</sub> activates PDE remains unknown however, it is speculated to be similar to the activation of PDE by GTP-bound T<sub>α</sub>.<ref name="pmid8143750"/>

In order to prevent activation of PDE in the dark, the concentration of GDP-bound T<sub>α</sub> should be kept to a minimum. This job seems to fall to the T<sub>βγ</sub> to keep the GDP-bound T<sub>α</sub> bound in the form of holotransducin.<ref name="pmid8143750"/>

For deactivation, hydrolysis of the bound GTP by the T<sub>α</sub> is necessary for T<sub>α</sub> deactivation and returning the transducin to its basal from. However, simple hydrolysis of GTP may not necessarily be enough to deactivate PDE. T<sub>βγ</sub> comes into play here again with an important role in PDE deactivation.<ref name="pmid8143750"/> The addition of T<sub>βγ</sub> facilitates inhibition of the PDE catalytic moiety because it binds with the T<sub>α</sub>-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<sub>α</sub>.<ref name="pmid2538446"/>

==Genes==
* Rods: GNAT1, GNB1, GNGT1; Cones: GNAT2, GNB3, GNGT2 <ref>{{Cite journal |last=Lamb |first=Trevor D. |date=September 2013 |title=Evolution of phototransduction, vertebrate photoreceptors and retina |url=https://linkinghub.elsevier.com/retrieve/pii/S1350946213000402 |journal=Progress in Retinal and Eye Research |language=en |volume=36 |pages=52–119 |doi=10.1016/j.preteyeres.2013.06.001|pmid=23792002 |s2cid=38219705 |hdl=1885/84715 |hdl-access=free }}</ref>

== References ==
{{reflist|2}}

==External links==
* {{MeshName|Transducin}}

{{Intracellular signaling peptides and proteins}}
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[[Category:G proteins]]