Sandbox Reserved 1170: Difference between revisions

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== Structure ==

[[Image:HGPR40bind2.png|200 px|right|thumb|Figure 1. Second proposed binding site of hGPR40 with surface shown. Substrate would bind in the deep pocket shown between TM3, 4, and 5.]]

Like most G-protein coupled receptors, hGPR40 contains <scene name='72/721541/Top_view_transmembrane_helices/2'>seven transmembrane helices</scene> (<scene name='72/721541/Top_view_transmembrane_helices/1'>top view of TM helices</scene>). To obtain a [https://en.wikipedia.org/wiki/Protein_crystallization crystallized structure] of the protein, four <scene name='72/721541/Stabilizing_mutations/4'>stabilizing mutations</scene> (<scene name='72/721541/L42a/3'>L42A</scene>, <scene name='72/721541/F88a/4'>F88A</scene>, <scene name='72/721541/G103a/3'>G103A</scene>, <scene name='72/721541/Y202f/3'>Y202F</scene>) were made to increase expression levels and thermal stability of the protein. These mutations did not significantly impact the enzyme's binding affinity with a known agonist, TAK-875.<ref name="Srivastava"/> A <scene name='72/721541/Lysozyme_crimson/2'>T4 Lysozyme</scene> (shown in <FONT COLOR="#DC143C">crimson</FONT>) was also added to intracellular loop 3 to aid in the formation of crystals. T4 Lysozyme also had little effect on TAK-875 binding.<ref name="Srivastava"/> For clarity, lysozyme is removed in all further renderings of hGPR40.

Like most G-protein coupled receptors, hGPR40 contains <scene name='72/721541/Top_view_transmembrane_helices/2'>seven transmembrane helices</scene> (<scene name='72/721541/Top_view_transmembrane_helices/1'>top view of TM helices</scene>). To obtain a [https://en.wikipedia.org/wiki/Protein_crystallization crystallized structure] of the protein, four <scene name='72/721541/Stabilizing_mutations/4'>stabilizing mutations</scene> (<scene name='72/721541/L42a/3'>L42A</scene>, <scene name='72/721541/F88a/4'>F88A</scene>, <scene name='72/721541/G103a/3'>G103A</scene>, <scene name='72/721541/Y202f/3'>Y202F</scene>) were made to increase expression levels and thermal stability of the protein. These mutations did not significantly impact the enzyme's binding affinity with a known agonist, TAK-875.<ref name="Srivastava"/> A <scene name='72/721541/Lysozyme_crimson/2'>T4 Lysozyme</scene> (shown in <FONT COLOR="#DC143C">crimson</FONT>) was also added to intracellular loop 3 to aid in the formation of crystals. T4 Lysozyme also had little effect on TAK-875 binding.<ref name="Srivastava"/> For clarity, lysozyme is removed in all further renderings of hGPR40. hGPR40 also contains an extracellular loop that is conserved among most G-protein coupled receptors (ECL2). This loop has two subsections and is involved in the permeability of the binding site.

=== Binding Sites ===

Sandbox Reserved 1170: Difference between revisions

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@@ Line 7: / Line 7: @@
 == Structure ==
 [[Image:HGPR40bind2.png|200 px|right|thumb|Figure 1. Second proposed binding site of hGPR40 with surface shown. Substrate would bind in the deep pocket shown between TM3, 4, and 5.]]
-Like most G-protein coupled receptors, hGPR40 contains <scene name='72/721541/Top_view_transmembrane_helices/2'>seven transmembrane helices</scene> (<scene name='72/721541/Top_view_transmembrane_helices/1'>top view of TM helices</scene>). To obtain a [https://en.wikipedia.org/wiki/Protein_crystallization crystallized structure] of the protein, four <scene name='72/721541/Stabilizing_mutations/4'>stabilizing mutations</scene> (<scene name='72/721541/L42a/3'>L42A</scene>, <scene name='72/721541/F88a/4'>F88A</scene>, <scene name='72/721541/G103a/3'>G103A</scene>, <scene name='72/721541/Y202f/3'>Y202F</scene>) were made to increase expression levels and thermal stability of the protein. These mutations did not significantly impact the enzyme's binding affinity with a known agonist, TAK-875.<ref name="Srivastava"/> A <scene name='72/721541/Lysozyme_crimson/2'>T4 Lysozyme</scene> (shown in <FONT COLOR="#DC143C">crimson</FONT>) was also added to intracellular loop 3 to aid in the formation of crystals. T4 Lysozyme also had little effect on TAK-875 binding.<ref name="Srivastava"/> For clarity, lysozyme is removed in all further renderings of hGPR40.
+Like most G-protein coupled receptors, hGPR40 contains <scene name='72/721541/Top_view_transmembrane_helices/2'>seven transmembrane helices</scene> (<scene name='72/721541/Top_view_transmembrane_helices/1'>top view of TM helices</scene>). To obtain a [https://en.wikipedia.org/wiki/Protein_crystallization crystallized structure] of the protein, four <scene name='72/721541/Stabilizing_mutations/4'>stabilizing mutations</scene> (<scene name='72/721541/L42a/3'>L42A</scene>, <scene name='72/721541/F88a/4'>F88A</scene>, <scene name='72/721541/G103a/3'>G103A</scene>, <scene name='72/721541/Y202f/3'>Y202F</scene>) were made to increase expression levels and thermal stability of the protein. These mutations did not significantly impact the enzyme's binding affinity with a known agonist, TAK-875.<ref name="Srivastava"/> A <scene name='72/721541/Lysozyme_crimson/2'>T4 Lysozyme</scene> (shown in <FONT COLOR="#DC143C">crimson</FONT>) was also added to intracellular loop 3 to aid in the formation of crystals. T4 Lysozyme also had little effect on TAK-875 binding.<ref name="Srivastava"/> For clarity, lysozyme is removed in all further renderings of hGPR40. hGPR40 also contains an extracellular loop that is conserved among most G-protein coupled receptors (ECL2). This loop has two subsections and is involved in the permeability of the binding site.
 === Binding Sites ===