Testgp - Revision history

Alexander Berchansky at 15:20, 25 February 2021

2021-02-25T15:20:24Z

← Older revision		Revision as of 15:20, 25 February 2021
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	== G-Protein Activation Cycle ==		== G-Protein Activation Cycle ==
	[[Image:Img.JPG\|~~600px~~\|G protein cycle for the β2AR–Gs complex. Reprinted by permission from Macmillan Publishers Ltd on behalf of Cancer Research UK: Nature 477, 549–555, copyright 2011]]		[[Image:Img.JPG\|400px\|G protein cycle for the β2AR–Gs complex. Reprinted by permission from Macmillan Publishers Ltd on behalf of Cancer Research UK: Nature 477, 549–555, copyright 2011]]

	The figure shows the G Protein cycle<ref>doi:10.1038/nature10361</ref> - an extracellular agonist binding to the β2AR leads to <scene name='70/701430/Receptor_morphing_animation/2'>conformational rearrangements</scene> of the cytoplasmic ends of transmembrane segments that enable the Gs heterotrimer to bind the receptor. GDP is released from the α subunit upon formation of β2AR–Gs complex. The GTP binds to the nucleotide-free α subunit resulting in dissociation of the α and βγ subunits from the receptor. The subunits regulate their respective effector proteins adenylyl cyclase (AC) and Ca2+ channels. The Gs heterotrimer reassembles from α and βγ subunits following hydrolysis of GTP to GDP in the α subunit.		The figure shows the G Protein cycle<ref>doi:10.1038/nature10361</ref> - an extracellular agonist binding to the β2AR leads to <scene name='70/701430/Receptor_morphing_animation/2'>conformational rearrangements</scene> of the cytoplasmic ends of transmembrane segments that enable the Gs heterotrimer to bind the receptor. GDP is released from the α subunit upon formation of β2AR–Gs complex. The GTP binds to the nucleotide-free α subunit resulting in dissociation of the α and βγ subunits from the receptor. The subunits regulate their respective effector proteins adenylyl cyclase (AC) and Ca2+ channels. The Gs heterotrimer reassembles from α and βγ subunits following hydrolysis of GTP to GDP in the α subunit.

Alexander Berchansky at 15:19, 25 February 2021

2021-02-25T15:19:48Z

← Older revision		Revision as of 15:19, 25 February 2021
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	{{Template:Button Toggle Animation2}}		{{Template:Button Toggle Animation2}}

	A previous structure of a GTPγS bound (i.e. active, "turned on") Gαs protein showed that both domains are involved in nucleotide binding, as the nucleotide-binding pocket of the Gαs subunit is formed by the interface between GαsRas and GαsAH<ref>doi:10.1126/science.278.5345.1943</ref>. It was also previously known that the GsαAH domain has a variable position relative to the GsαRas domain between this GTP bound (active) state and the nucleotide free state<ref>DOI:10.1126/science.8266082</ref><ref>doi:10.1073/pnas.1105810108</ref><ref>doi:10.1073/pnas.1113645108</ref><ref>doi:10.1038/nature10488</ref>. However, the β2AR–Gs complex structure of the receptor attached to the empty (no guanosine phosphate attached) G protein enabled comparing it to the active (GTP bound) structure and by that showing <scene name='70/701430/Gamorph/2'>how large this displacement is</scene> - this is probably the most surprising observation arising from the β2AR–Gs complex.

	~~{{Template:Button Toggle Animation2}}~~

	==See Also==		==See Also==

Alexander Berchansky at 15:19, 25 February 2021

2021-02-25T15:19:12Z

@@ Line 21: / Line 21: @@
 == G-Protein Activation Cycle ==
-[[Image:Img.JPG|450px|G protein cycle for the β2AR–Gs complex. Reprinted by permission from Macmillan Publishers Ltd on behalf of Cancer Research UK: Nature 477, 549–555, copyright 2011]]
+[[Image:Img.JPG|600px|G protein cycle for the β2AR–Gs complex. Reprinted by permission from Macmillan Publishers Ltd on behalf of Cancer Research UK: Nature 477, 549–555, copyright 2011]]
 The figure shows the G Protein cycle<ref>doi:10.1038/nature10361</ref> - an extracellular agonist binding to the β2AR leads to <scene name='70/701430/Receptor_morphing_animation/2'>conformational rearrangements</scene> of the cytoplasmic ends of transmembrane segments that enable the Gs heterotrimer to bind the receptor. GDP is released from the α subunit upon formation of β2AR–Gs complex. The GTP binds to the nucleotide-free α subunit resulting in dissociation of the α and βγ subunits from the receptor. The subunits regulate their respective effector proteins adenylyl cyclase (AC) and Ca2+ channels. The Gs heterotrimer reassembles from α and βγ subunits following hydrolysis of GTP to GDP in the α subunit.
 {{Template:Button Toggle Animation2}}

Alexander Berchansky at 15:16, 25 February 2021

2021-02-25T15:16:32Z

← Older revision		Revision as of 15:16, 25 February 2021
Line 21:		Line 21:

	== G-Protein Activation Cycle ==		== G-Protein Activation Cycle ==
	[[Image:Img.JPG\|~~600px~~\|G protein cycle for the β2AR–Gs complex. Reprinted by permission from Macmillan Publishers Ltd on behalf of Cancer Research UK: Nature 477, 549–555, copyright 2011]]		[[Image:Img.JPG\|450px\|G protein cycle for the β2AR–Gs complex. Reprinted by permission from Macmillan Publishers Ltd on behalf of Cancer Research UK: Nature 477, 549–555, copyright 2011]]

	The figure shows the G Protein cycle<ref>doi:10.1038/nature10361</ref> - an extracellular agonist binding to the β2AR leads to <scene name='70/701430/Receptor_morphing_animation/2'>conformational rearrangements</scene> of the cytoplasmic ends of transmembrane segments that enable the Gs heterotrimer to bind the receptor. GDP is released from the α subunit upon formation of β2AR–Gs complex. The GTP binds to the nucleotide-free α subunit resulting in dissociation of the α and βγ subunits from the receptor. The subunits regulate their respective effector proteins adenylyl cyclase (AC) and Ca2+ channels. The Gs heterotrimer reassembles from α and βγ subunits following hydrolysis of GTP to GDP in the α subunit.		The figure shows the G Protein cycle<ref>doi:10.1038/nature10361</ref> - an extracellular agonist binding to the β2AR leads to <scene name='70/701430/Receptor_morphing_animation/2'>conformational rearrangements</scene> of the cytoplasmic ends of transmembrane segments that enable the Gs heterotrimer to bind the receptor. GDP is released from the α subunit upon formation of β2AR–Gs complex. The GTP binds to the nucleotide-free α subunit resulting in dissociation of the α and βγ subunits from the receptor. The subunits regulate their respective effector proteins adenylyl cyclase (AC) and Ca2+ channels. The Gs heterotrimer reassembles from α and βγ subunits following hydrolysis of GTP to GDP in the α subunit.

	{{Template:Button Toggle Animation2}}		{{Template:Button Toggle Animation2}}

Alexander Berchansky at 15:14, 25 February 2021

2021-02-25T15:14:15Z

← Older revision		Revision as of 15:14, 25 February 2021
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	The figure shows the G Protein cycle<ref>doi:10.1038/nature10361</ref> - an extracellular agonist binding to the β2AR leads to <scene name='70/701430/Receptor_morphing_animation/2'>conformational rearrangements</scene> of the cytoplasmic ends of transmembrane segments that enable the Gs heterotrimer to bind the receptor. GDP is released from the α subunit upon formation of β2AR–Gs complex. The GTP binds to the nucleotide-free α subunit resulting in dissociation of the α and βγ subunits from the receptor. The subunits regulate their respective effector proteins adenylyl cyclase (AC) and Ca2+ channels. The Gs heterotrimer reassembles from α and βγ subunits following hydrolysis of GTP to GDP in the α subunit.		The figure shows the G Protein cycle<ref>doi:10.1038/nature10361</ref> - an extracellular agonist binding to the β2AR leads to <scene name='70/701430/Receptor_morphing_animation/2'>conformational rearrangements</scene> of the cytoplasmic ends of transmembrane segments that enable the Gs heterotrimer to bind the receptor. GDP is released from the α subunit upon formation of β2AR–Gs complex. The GTP binds to the nucleotide-free α subunit resulting in dissociation of the α and βγ subunits from the receptor. The subunits regulate their respective effector proteins adenylyl cyclase (AC) and Ca2+ channels. The Gs heterotrimer reassembles from α and βγ subunits following hydrolysis of GTP to GDP in the α subunit.

	{{Template:Button Toggle Animation2}}		{{Template:Button Toggle Animation2}}

	~~== G-Protein variability ==~~
	The Gαs subunit consists of two domains, the <scene name='70/701430/Alpharas/2'>Ras domain (GαsRas)</scene> and the <scene name='70/701430/Alphahelical/2'>α-helical domain (GαsAH)</scene>. A previous structure of a GTPγS bound (i.e. active, "turned on") Gαs protein (PDB ID: [http://www.rcsb.org/pdb/explore/explore.do?structureId=1AZT 1AZT]) showed that both domains are involved in nucleotide binding, as the nucleotide-binding pocket of the Gαs subunit is formed by the interface between GαsRas and GαsAH<ref>doi:10.1126/science.278.5345.1943</ref>. It was also previously known that the GsαAH domain has a variable position relative to the GsαRas domain between this GTP bound (active) state and the nucleotide free state<ref>DOI:10.1126/science.8266082</ref><ref>doi:10.1073/pnas.1105810108</ref><ref>doi:10.1073/pnas.1113645108</ref><ref>doi:10.1038/nature10488</ref>. However, the β2AR–Gs complex structure of the receptor attached to the empty (no guanosine phosphate attached) G protein enabled comparing it to the active (GTP bound) structure and by that showing <scene name='70/701430/Gamorph/2'>how large this displacement is</scene> - this is probably the most surprising observation arising from the β2AR–Gs complex.

Alexander Berchansky at 15:13, 25 February 2021

2021-02-25T15:13:55Z

← Older revision		Revision as of 15:13, 25 February 2021
Line 29:		Line 29:
	== G-Protein variability ==		== G-Protein variability ==
	The Gαs subunit consists of two domains, the <scene name='70/701430/Alpharas/2'>Ras domain (GαsRas)</scene> and the <scene name='70/701430/Alphahelical/2'>α-helical domain (GαsAH)</scene>. A previous structure of a GTPγS bound (i.e. active, "turned on") Gαs protein (PDB ID: [http://www.rcsb.org/pdb/explore/explore.do?structureId=1AZT 1AZT]) showed that both domains are involved in nucleotide binding, as the nucleotide-binding pocket of the Gαs subunit is formed by the interface between GαsRas and GαsAH<ref>doi:10.1126/science.278.5345.1943</ref>. It was also previously known that the GsαAH domain has a variable position relative to the GsαRas domain between this GTP bound (active) state and the nucleotide free state<ref>DOI:10.1126/science.8266082</ref><ref>doi:10.1073/pnas.1105810108</ref><ref>doi:10.1073/pnas.1113645108</ref><ref>doi:10.1038/nature10488</ref>. However, the β2AR–Gs complex structure of the receptor attached to the empty (no guanosine phosphate attached) G protein enabled comparing it to the active (GTP bound) structure and by that showing <scene name='70/701430/Gamorph/2'>how large this displacement is</scene> - this is probably the most surprising observation arising from the β2AR–Gs complex.		The Gαs subunit consists of two domains, the <scene name='70/701430/Alpharas/2'>Ras domain (GαsRas)</scene> and the <scene name='70/701430/Alphahelical/2'>α-helical domain (GαsAH)</scene>. A previous structure of a GTPγS bound (i.e. active, "turned on") Gαs protein (PDB ID: [http://www.rcsb.org/pdb/explore/explore.do?structureId=1AZT 1AZT]) showed that both domains are involved in nucleotide binding, as the nucleotide-binding pocket of the Gαs subunit is formed by the interface between GαsRas and GαsAH<ref>doi:10.1126/science.278.5345.1943</ref>. It was also previously known that the GsαAH domain has a variable position relative to the GsαRas domain between this GTP bound (active) state and the nucleotide free state<ref>DOI:10.1126/science.8266082</ref><ref>doi:10.1073/pnas.1105810108</ref><ref>doi:10.1073/pnas.1113645108</ref><ref>doi:10.1038/nature10488</ref>. However, the β2AR–Gs complex structure of the receptor attached to the empty (no guanosine phosphate attached) G protein enabled comparing it to the active (GTP bound) structure and by that showing <scene name='70/701430/Gamorph/2'>how large this displacement is</scene> - this is probably the most surprising observation arising from the β2AR–Gs complex.

	~~{{Template:Button Toggle Animation2}}~~

Alexander Berchansky at 15:13, 25 February 2021

2021-02-25T15:13:38Z

← Older revision		Revision as of 15:13, 25 February 2021
Line 25:		Line 25:
	The figure shows the G Protein cycle<ref>doi:10.1038/nature10361</ref> - an extracellular agonist binding to the β2AR leads to <scene name='70/701430/Receptor_morphing_animation/2'>conformational rearrangements</scene> of the cytoplasmic ends of transmembrane segments that enable the Gs heterotrimer to bind the receptor. GDP is released from the α subunit upon formation of β2AR–Gs complex. The GTP binds to the nucleotide-free α subunit resulting in dissociation of the α and βγ subunits from the receptor. The subunits regulate their respective effector proteins adenylyl cyclase (AC) and Ca2+ channels. The Gs heterotrimer reassembles from α and βγ subunits following hydrolysis of GTP to GDP in the α subunit.		The figure shows the G Protein cycle<ref>doi:10.1038/nature10361</ref> - an extracellular agonist binding to the β2AR leads to <scene name='70/701430/Receptor_morphing_animation/2'>conformational rearrangements</scene> of the cytoplasmic ends of transmembrane segments that enable the Gs heterotrimer to bind the receptor. GDP is released from the α subunit upon formation of β2AR–Gs complex. The GTP binds to the nucleotide-free α subunit resulting in dissociation of the α and βγ subunits from the receptor. The subunits regulate their respective effector proteins adenylyl cyclase (AC) and Ca2+ channels. The Gs heterotrimer reassembles from α and βγ subunits following hydrolysis of GTP to GDP in the α subunit.

			{{Template:Button Toggle Animation2}}

			== G-Protein variability ==
			The Gαs subunit consists of two domains, the <scene name='70/701430/Alpharas/2'>Ras domain (GαsRas)</scene> and the <scene name='70/701430/Alphahelical/2'>α-helical domain (GαsAH)</scene>. A previous structure of a GTPγS bound (i.e. active, "turned on") Gαs protein (PDB ID: [http://www.rcsb.org/pdb/explore/explore.do?structureId=1AZT 1AZT]) showed that both domains are involved in nucleotide binding, as the nucleotide-binding pocket of the Gαs subunit is formed by the interface between GαsRas and GαsAH<ref>doi:10.1126/science.278.5345.1943</ref>. It was also previously known that the GsαAH domain has a variable position relative to the GsαRas domain between this GTP bound (active) state and the nucleotide free state<ref>DOI:10.1126/science.8266082</ref><ref>doi:10.1073/pnas.1105810108</ref><ref>doi:10.1073/pnas.1113645108</ref><ref>doi:10.1038/nature10488</ref>. However, the β2AR–Gs complex structure of the receptor attached to the empty (no guanosine phosphate attached) G protein enabled comparing it to the active (GTP bound) structure and by that showing <scene name='70/701430/Gamorph/2'>how large this displacement is</scene> - this is probably the most surprising observation arising from the β2AR–Gs complex.

	{{Template:Button Toggle Animation2}}		{{Template:Button Toggle Animation2}}

Alexander Berchansky at 15:13, 25 February 2021

2021-02-25T15:13:18Z

← Older revision		Revision as of 15:13, 25 February 2021
Line 23:		Line 23:
	[[Image:Img.JPG\|600px\|G protein cycle for the β2AR–Gs complex. Reprinted by permission from Macmillan Publishers Ltd on behalf of Cancer Research UK: Nature 477, 549–555, copyright 2011]]		[[Image:Img.JPG\|600px\|G protein cycle for the β2AR–Gs complex. Reprinted by permission from Macmillan Publishers Ltd on behalf of Cancer Research UK: Nature 477, 549–555, copyright 2011]]

	The figure shows the G Protein cycle<ref>doi:10.1038/nature10361</ref> - an extracellular agonist binding to the β2AR leads to <scene name='70/701430/Receptor_morphing_animation/2'>conformational rearrangements</scene> of the cytoplasmic ends of transmembrane segments that enable the Gs heterotrimer to bind the receptor. GDP is released from the α subunit upon formation of β2AR–Gs complex. The GTP binds to the nucleotide-free α subunit resulting in dissociation of the α and βγ subunits from the receptor. The subunits regulate their respective effector proteins adenylyl cyclase (AC) and Ca2+ channels. The Gs heterotrimer reassembles from α and βγ subunits following hydrolysis of GTP to GDP in the α subunit.		The figure shows the G Protein cycle<ref>doi:10.1038/nature10361</ref> - an extracellular agonist binding to the β2AR leads to <scene name='70/701430/Receptor_morphing_animation/2'>conformational rearrangements</scene> of the cytoplasmic ends of transmembrane segments that enable the Gs heterotrimer to bind the receptor. GDP is released from the α subunit upon formation of β2AR–Gs complex. The GTP binds to the nucleotide-free α subunit resulting in dissociation of the α and βγ subunits from the receptor. The subunits regulate their respective effector proteins adenylyl cyclase (AC) and Ca2+ channels. The Gs heterotrimer reassembles from α and βγ subunits following hydrolysis of GTP to GDP in the α subunit.

			{{Template:Button Toggle Animation2}}

Alexander Berchansky at 15:12, 25 February 2021

2021-02-25T15:12:42Z

← Older revision		Revision as of 15:12, 25 February 2021
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	<StructureSection load='3sn6' size='340' side='right' caption='Adrenergic receptor (blue) complex with G protein α subunit (grey), β subunit (green), γ-2 subunit (gold), antibody fragment (Orchid) and benzoxazin derivative [[3sn6]], [[Resolution\|resolution]] 3.20Å' scene=''>		<StructureSection load='3sn6' size='340' side='right' caption='Adrenergic receptor (blue) complex with G protein α subunit (grey), β subunit (green), γ-2 subunit (gold), antibody fragment (Orchid) and benzoxazin derivative [[3sn6]], [[Resolution\|resolution]] 3.20Å' scene=''>

			==Introduction==
			[[G protein-coupled receptors]] (GPCRs) are a large family of protein receptors, which have seven-transmembrane helices and are found over a large array of eukaryotic cells. These receptors take a major part in a multitude of signal transduction pathways, including amongst others responses to hormones and neurotransmitters, sensing light, taste and smell, and many more. These receptors are also involved in many different types of diseases and are the target of almost 50% of current medical drugs.
			The [[Beta-2 Adrenergic Receptors]] are a type of GPCR that are activated by catecholamine hormone ligands such as adrenaline (epinephrine). These receptors are responsible for many of the adrenaline related (“fight-or-flight”) responses and functions, and are used as a common model system for the GPCR family.
			GPCRs bind their ligand and [[Group:SMART:A Physical Model of the β2-Adrenergic Receptor\|overcome a conformational change]] that activates an attached [[Guanine nucleotide-binding protein]] (G protein) and allows it to detach from the cellular end of the receptor and start the different signal transduction pathways.

			G proteins<ref>https://en.wikipedia.org/wiki/G_protein</ref> are a family of proteins that act as molecular switches inside cells. G proteins belong to the larger group of enzymes called GTPases<ref>https://en.wikipedia.org/wiki/GTPase</ref>, and appear either as monomeric small GTPases<ref>https://en.wikipedia.org/wiki/Small_GTPase</ref>, or as heterotrimeric G protein complexes<ref>https://en.wikipedia.org/wiki/Heterotrimeric_G_protein</ref> that are made up of alpha (α), beta (β) and gamma (γ) subunits<ref>doi:10.1093/dnares/7.2.111</ref>. When they are bound to guanosine triphosphate ([https://en.wikipedia.org/wiki/Guanosine_triphosphate GTP]), they are 'on', and when they are bound to guanosine diphosphate ([https://en.wikipedia.org/wiki/Guanosine_diphosphate GDP]), they are 'off'.

			Since these receptors have seven transmembrane helices as well as inner and outer cell regions, they are very difficult to purify and crystallize. Some crystal structures have been determined for the inactive receptors as well as for the G proteins that they bind. PDB entry 3SN6 is the first structure of the full complex of the Beta 2 Adrenergic Receptor bound to Gs in their active state, and it provides the first high-resolution insight into the mechanism of signal transduction across the plasma membrane by a GPCR.

			== Complex structure ==
			The overall structure shows the <scene name='70/701430/Receptor/1'>β2AR receptor</scene> (dark blue) bound to an agonist (in spheres) along with a <scene name='70/701430/Lysozyme/1'>T4</scene> [https://en.wikipedia.org/wiki/Lysozyme lysozyme] fused to its amino terminus in order to facilitate crystallization. The receptor interacts with <scene name='70/701430/Galpha/1'>Gαs</scene> (light blue). Gαs together with <scene name='70/701430/Gbeta/1'>Gβ</scene> (light green) and <scene name='70/701430/Ggamma/1'>Gγ</scene> (gold) constitute the heterotrimeric G protein Gs. A Gs-binding [https://en.wikipedia.org/wiki/Single-domain_antibody nanobody] <scene name='70/701430/Nanobody/1'>(in pink)</scene>, which also facilitates crystallization, binds the G protein between the α and β subunits.

			{{Template:Button Toggle Animation2}}

			== G-Protein-GPCR Interactions ==
			The α5-helix of Gαs <scene name='70/701430/Docking/2'>docks</scene> into a cavity formed on the intracellular side of the receptor by the opening of the transmembrane helices. Within the transmembrane core, the interactions are primarily non-polar - an exception involves <scene name='70/701430/Receptor_g_protein_interaction/4'>packing</scene> of Tyr 391 of the α5-helix against Arg 131 of the conserved DRY sequence in TM3. Arg 131 also packs against Tyr 326 of the conserved NPxxY sequence in TM7. As the α5-helix exits the receptor it forms a network of polar interactions with TM5 and TM3. Receptor residues Thr 68 and Asp 130 interact with the ICL2 helix of the β2AR via Tyr 141, positioning the helix so that Phe 139 of the receptor <scene name='70/701430/Receptor_gprotein_interaction2/1'>docks into</scene> a hydrophobic pocket on the G protein surface, thereby structurally linking receptor–G protein interactions with the highly conserved DRY motif of the β2AR.

			{{Template:Button Toggle Animation2}}

			== G-Protein Activation Cycle ==
			[[Image:Img.JPG\|600px\|G protein cycle for the β2AR–Gs complex. Reprinted by permission from Macmillan Publishers Ltd on behalf of Cancer Research UK: Nature 477, 549–555, copyright 2011]]

			The figure shows the G Protein cycle<ref>doi:10.1038/nature10361</ref> - an extracellular agonist binding to the β2AR leads to <scene name='70/701430/Receptor_morphing_animation/2'>conformational rearrangements</scene> of the cytoplasmic ends of transmembrane segments that enable the Gs heterotrimer to bind the receptor. GDP is released from the α subunit upon formation of β2AR–Gs complex. The GTP binds to the nucleotide-free α subunit resulting in dissociation of the α and βγ subunits from the receptor. The subunits regulate their respective effector proteins adenylyl cyclase (AC) and Ca2+ channels. The Gs heterotrimer reassembles from α and βγ subunits following hydrolysis of GTP to GDP in the α subunit.

Alexander Berchansky: Replacing page with '
2021-02-25T15:12:17Z

Replacing page with '<StructureSection load='3sn6' size='340' side='right' caption='Adrenergic receptor (blue) complex with G protein α subunit (grey), β subunit (green), γ-2 subunit (gold), ant...'

← Older revision Revision as of 15:12, 25 February 2021
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<StructureSection load='3sn6' size='340' side='right' caption='Adrenergic receptor (blue) complex with G protein α subunit (grey), β subunit (green), γ-2 subunit (gold), antibody fragment (Orchid) and benzoxazin derivative [[3sn6]], [[Resolution|resolution]] 3.20Å' scene=''>
<StructureSection load='3sn6' size='340' side='right' caption='Adrenergic receptor (blue) complex with G protein α subunit (grey), β subunit (green), γ-2 subunit (gold), antibody fragment (Orchid) and benzoxazin derivative [[3sn6]], [[Resolution|resolution]] 3.20Å' scene=''>

==Introduction==

[[G protein-coupled receptors]] (GPCRs) are a large family of protein receptors, which have seven-transmembrane helices and are found over a large array of eukaryotic cells. These receptors take a major part in a multitude of signal transduction pathways, including amongst others responses to hormones and neurotransmitters, sensing light, taste and smell, and many more. These receptors are also involved in many different types of diseases and are the target of almost 50% of current medical drugs.

The [[Beta-2 Adrenergic Receptors]] are a type of GPCR that are activated by catecholamine hormone ligands such as adrenaline (epinephrine). These receptors are responsible for many of the adrenaline related (“fight-or-flight”) responses and functions, and are used as a common model system for the GPCR family.

GPCRs bind their ligand and [[Group:SMART:A Physical Model of the β2-Adrenergic Receptor|overcome a conformational change]] that activates an attached [[Guanine nucleotide-binding protein]] (G protein) and allows it to detach from the cellular end of the receptor and start the different signal transduction pathways.

G proteins<ref>https://en.wikipedia.org/wiki/G_protein</ref> are a family of proteins that act as molecular switches inside cells. G proteins belong to the larger group of enzymes called GTPases<ref>https://en.wikipedia.org/wiki/GTPase</ref>, and appear either as monomeric small GTPases<ref>https://en.wikipedia.org/wiki/Small_GTPase</ref>, or as heterotrimeric G protein complexes<ref>https://en.wikipedia.org/wiki/Heterotrimeric_G_protein</ref> that are made up of alpha (α), beta (β) and gamma (γ) subunits<ref>doi:10.1093/dnares/7.2.111</ref>. When they are bound to guanosine triphosphate ([https://en.wikipedia.org/wiki/Guanosine_triphosphate GTP]), they are 'on', and when they are bound to guanosine diphosphate ([https://en.wikipedia.org/wiki/Guanosine_diphosphate GDP]), they are 'off'.

Since these receptors have seven transmembrane helices as well as inner and outer cell regions, they are very difficult to purify and crystallize. Some crystal structures have been determined for the inactive receptors as well as for the G proteins that they bind. PDB entry 3SN6 is the first structure of the full complex of the Beta 2 Adrenergic Receptor bound to Gs in their active state, and it provides the first high-resolution insight into the mechanism of signal transduction across the plasma membrane by a GPCR.

== Complex structure ==

The overall structure shows the <scene name='70/701430/Receptor/1'>β2AR receptor</scene> (dark blue) bound to an agonist (in spheres) along with a <scene name='70/701430/Lysozyme/1'>T4</scene> [https://en.wikipedia.org/wiki/Lysozyme lysozyme] fused to its amino terminus in order to facilitate crystallization. The receptor interacts with <scene name='70/701430/Galpha/1'>Gαs</scene> (light blue). Gαs together with <scene name='70/701430/Gbeta/1'>Gβ</scene> (light green) and <scene name='70/701430/Ggamma/1'>Gγ</scene> (gold) constitute the heterotrimeric G protein Gs. A Gs-binding [https://en.wikipedia.org/wiki/Single-domain_antibody nanobody] <scene name='70/701430/Nanobody/1'>(in pink)</scene>, which also facilitates crystallization, binds the G protein between the α and β subunits.

{{Template:Button Toggle Animation2}}

== G-Protein-GPCR Interactions ==

The α5-helix of Gαs <scene name='70/701430/Docking/2'>docks</scene> into a cavity formed on the intracellular side of the receptor by the opening of the transmembrane helices. Within the transmembrane core, the interactions are primarily non-polar - an exception involves <scene name='70/701430/Receptor_g_protein_interaction/4'>packing</scene> of Tyr 391 of the α5-helix against Arg 131 of the conserved DRY sequence in TM3. Arg 131 also packs against Tyr 326 of the conserved NPxxY sequence in TM7. As the α5-helix exits the receptor it forms a network of polar interactions with TM5 and TM3. Receptor residues Thr 68 and Asp 130 interact with the ICL2 helix of the β2AR via Tyr 141, positioning the helix so that Phe 139 of the receptor <scene name='70/701430/Receptor_gprotein_interaction2/1'>docks into</scene> a hydrophobic pocket on the G protein surface, thereby structurally linking receptor–G protein interactions with the highly conserved DRY motif of the β2AR.

{{Template:Button Toggle Animation2}}

== G-Protein Activation Cycle ==

[[Image:Img.JPG|600px|G protein cycle for the β2AR–Gs complex. Reprinted by permission from Macmillan Publishers Ltd on behalf of Cancer Research UK: Nature 477, 549–555, copyright 2011]]

The figure shows the G Protein cycle<ref>doi:10.1038/nature10361</ref> - an extracellular agonist binding to the β2AR leads to <scene name='70/701430/Receptor_morphing_animation/2'>conformational rearrangements</scene> of the cytoplasmic ends of transmembrane segments that enable the Gs heterotrimer to bind the receptor. GDP is released from the α subunit upon formation of β2AR–Gs complex. The GTP binds to the nucleotide-free α subunit resulting in dissociation of the α and βγ subunits from the receptor. The subunits regulate their respective effector proteins adenylyl cyclase (AC) and Ca2+ channels. The Gs heterotrimer reassembles from α and βγ subunits following hydrolysis of GTP to GDP in the α subunit.

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== G-Protein variability ==

The Gαs subunit consists of two domains, the <scene name='70/701430/Alpharas/2'>Ras domain (GαsRas)</scene> and the <scene name='70/701430/Alphahelical/2'>α-helical domain (GαsAH)</scene>. A previous structure of a GTPγS bound (i.e. active, "turned on") Gαs protein (PDB ID: [http://www.rcsb.org/pdb/explore/explore.do?structureId=1AZT 1AZT]) showed that both domains are involved in nucleotide binding, as the nucleotide-binding pocket of the Gαs subunit is formed by the interface between GαsRas and GαsAH<ref>doi:10.1126/science.278.5345.1943</ref>. It was also previously known that the GsαAH domain has a variable position relative to the GsαRas domain between this GTP bound (active) state and the nucleotide free state<ref>DOI:10.1126/science.8266082</ref><ref>doi:10.1073/pnas.1105810108</ref><ref>doi:10.1073/pnas.1113645108</ref><ref>doi:10.1038/nature10488</ref>. However, the β2AR–Gs complex structure of the receptor attached to the empty (no guanosine phosphate attached) G protein enabled comparing it to the active (GTP bound) structure and by that showing <scene name='70/701430/Gamorph/2'>how large this displacement is</scene> - this is probably the most surprising observation arising from the β2AR–Gs complex.