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From Proteopedia
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Like most G-protein coupled receptors, hGPR40 contains '''seven transmembrane helices'''. In order obtain a crystallized structure of the protein, four <scene name='72/721541/Stabilizing_mutations/3'>stabilizing mutations</scene> (<scene name='72/721541/L42a/2'>L42A</scene>, <scene name='72/721541/F88a/2'>F88A</scene>, <scene name='72/721541/G103a/2'>G103A</scene>, <scene name='72/721541/Y202f/2'>Y202F</scene>) were made to increase expression levels and thermal stability of the protein.<ref name="Srivastava"/> | Like most G-protein coupled receptors, hGPR40 contains '''seven transmembrane helices'''. In order obtain a crystallized structure of the protein, four <scene name='72/721541/Stabilizing_mutations/3'>stabilizing mutations</scene> (<scene name='72/721541/L42a/2'>L42A</scene>, <scene name='72/721541/F88a/2'>F88A</scene>, <scene name='72/721541/G103a/2'>G103A</scene>, <scene name='72/721541/Y202f/2'>Y202F</scene>) were made to increase expression levels and thermal stability of the protein.<ref name="Srivastava"/> | ||
=== Charge Network === | === Charge Network === | ||
| - | hGPR40 has a distinct binding pocket that is established by | + | hGPR40 has a distinct binding pocket that is established by seven key residues. The importance of these residues for agonist binding was determined by mutagenesis studies. Each of these residues have either a charged or polar R-group that allows them to develop a charge network that interacts with the carboxylate moiety of agonists. In 2007 and 2009 researchers showed the presence of Arg 183 and Arg 258 in the binding pocket <ref name="Sum">PMID: 17699519</ref><ref name="Sum, C.">PMID:19068482</ref> Along with the two Arginine residues, the charge network incorporates two Tyrosine residues.These residues (Tyr 91 and Tyr 240) also stabilize the carboxylate of the agonists. It was further determined that Tyr 240 is epecially important for binding. Mutation of Tyr 240 caused a reduction in the binding affinity of TAK-875 by eight fold and had a significant effect on the Kd of the protein.<ref name="Srivastava"/> |
[[Image:charge network residues.png|300 px|right|thumb|Figure Legend]] | [[Image:charge network residues.png|300 px|right|thumb|Figure Legend]] | ||
<scene name='72/721541/Hydrogen_binding_1/3'>Key Binding Residues</scene> | <scene name='72/721541/Hydrogen_binding_1/3'>Key Binding Residues</scene> | ||
Revision as of 18:55, 28 March 2016
Human GPR40
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References
- ↑ Hanson, R. M., Prilusky, J., Renjian, Z., Nakane, T. and Sussman, J. L. (2013), JSmol and the Next-Generation Web-Based Representation of 3D Molecular Structure as Applied to Proteopedia. Isr. J. Chem., 53:207-216. doi:http://dx.doi.org/10.1002/ijch.201300024
- ↑ Herraez A. Biomolecules in the computer: Jmol to the rescue. Biochem Mol Biol Educ. 2006 Jul;34(4):255-61. doi: 10.1002/bmb.2006.494034042644. PMID:21638687 doi:10.1002/bmb.2006.494034042644
- ↑ 3.0 3.1 3.2 Srivastava A, Yano J, Hirozane Y, Kefala G, Gruswitz F, Snell G, Lane W, Ivetac A, Aertgeerts K, Nguyen J, Jennings A, Okada K. High-resolution structure of the human GPR40 receptor bound to allosteric agonist TAK-875. Nature. 2014 Jul 20. doi: 10.1038/nature13494. PMID:25043059 doi:http://dx.doi.org/10.1038/nature13494
- ↑ Sum CS, Tikhonova IG, Neumann S, Engel S, Raaka BM, Costanzi S, Gershengorn MC. Identification of residues important for agonist recognition and activation in GPR40. J Biol Chem. 2007 Oct 5;282(40):29248-55. Epub 2007 Aug 15. PMID:17699519 doi:http://dx.doi.org/10.1074/jbc.M705077200
- ↑ Sum CS, Tikhonova IG, Costanzi S, Gershengorn MC. Two arginine-glutamate ionic locks near the extracellular surface of FFAR1 gate receptor activation. J Biol Chem. 2009 Feb 6;284(6):3529-36. doi: 10.1074/jbc.M806987200. Epub 2008, Dec 8. PMID:19068482 doi:http://dx.doi.org/10.1074/jbc.M806987200
