{"@context":"http://iiif.io/api/presentation/2/context.json","@id":"https://repo.library.stonybrook.edu/cantaloupe/iiif/2/manifest.json","@type":"sc:Manifest","label":"Correlating Structure and Function in Class A \nGPCRs","metadata":[{"label":"dc.description.sponsorship","value":"This work is sponsored by the Stony Brook University Graduate School in compliance with the requirements for completion of degree."},{"label":"dc.format","value":"Monograph"},{"label":"dc.format.medium","value":"Electronic Resource"},{"label":"dc.identifier.uri","value":"http://hdl.handle.net/11401/71237"},{"label":"dc.language.iso","value":"en_US"},{"label":"dc.publisher","value":"The Graduate School, Stony Brook University: Stony Brook, NY."},{"label":"dcterms.abstract","value":"Class A \nG protein-coupled receptors (GPCRs) serve as the gatekeepers for cell signaling in eukaryotes. With over 4% of the \nhuman protein-encoding genome dedicated to their expression, GPCRs are accountable for a variety of physiological \nresponses including, vision, vasodilation, and cell migration. These receptors all contain seven transmembrane \nhelices and a number of conserved residues suggesting a universal activation mechanism. In order to understand GPCR \nactivation, it is essential to delineate the structural differences between ligand-bound receptor conformations. \nDespite the breadth of biophysical studies conducted to date, how ligand binding is coupled to receptor activation \nremains to be elucidated. In this thesis, solid-state NMR studies are presented that target conformational changes in \nthe low light visual pigment rhodopsin, a prototypical GPCR. Rhodopsin is activated by a light-induced 11-cis to \ntrans isomerization of a covalently bound retinal chromophore. The experimental data presented define global \nstructural changes that couple receptor activation with the binding of downstream signaling targets. \nActivation-induced changes are described in the region of transmembrane helices H5 and H6. First, NMR distances \nmeasurements are used to temporally separate the motion of H6. Using 13C...13C dipolar couplings we observe a \nrotation of transmembrane helix H6 upon formation of Meta I. Meta I is the inactive predecessor of the signaling \ncompetent state, Meta II. Rotation of H6 in Meta I reflects the disruption of a salt bridge between \nArg135<super>3.50</super> and Glu247<super>6.40</super> prior to displacement of the \ntransmembrane helix in Meta II, which is required for coupling to heterotrimeric G protein. In addition, we show that \nH5 undergoes a rotation in the transition to Meta II. Specifically, we observe NMR contacts between \nTyr223<super>5.58</super>, Tyr306<super>7.53</super>, Met257<super>6.40</super>, \nand Arg135<super>3.50</super> that reveal a close association between these residues in the active Meta \nII state. Rotation of H5 allows a direct interaction to form between signature-conserved residues \nTyr223<super>5.58</super> and Arg135<super>3.50</super>. Fluorescence spectroscopy is used to \nmeasure the rate of active state decay. We find that the Tyr223<super>5.58</super> and \nArg135<super>3.50</super> interaction is crucial in stabilizing the active conformation of H5. The \nstructural studies on rhodopsin are extended to the ligand-activated \u0392#60;sub>2</sub>-adrenergic \nreceptor. We use NMR to probe the rotational orientation of transmembrane helix H5 in the presence of various \nligands. The data show a graded rotation of H5 that correlates with ligand efficacy.  Together, the structural \nstudies on rhodopsin and the \u0392<sub>2</sub>-adrenergic receptor reveal that H5 rotation is a \ncommon \nelement of GPCR activation."},{"label":"dcterms.available","value":"2015-04-24T14:46:37Z"},{"label":"dcterms.contributor","value":"Smith, Steven O, Scarlata, \nSuzanne"},{"label":"dcterms.creator","value":"Goncalves, Joseph Anthony"},{"label":"dcterms.dateAccepted","value":"2013-05-22T17:34:36Z"},{"label":"dcterms.dateSubmitted","value":"2015-04-24T14:46:37Z"},{"label":"dcterms.description","value":"Department of Molecular and \nCellular Biology"},{"label":"dcterms.extent","value":"149 \npg."},{"label":"dcterms.format","value":"Application/PDF"},{"label":"dcterms.identifier","value":"http://hdl.handle.net/1951/59665"},{"label":"dcterms.issued","value":"2012-08-01"},{"label":"dcterms.language","value":"en_US"},{"label":"dcterms.provenance","value":"Made available in DSpace on 2015-04-24T14:46:37Z (GMT). No. of bitstreams: 3\nGoncalves_grad.sunysb_0771E_11045.pdf.jpg: 1894 bytes, checksum: a6009c46e6ec8251b348085684cba80d (MD5)\nGoncalves_grad.sunysb_0771E_11045.pdf.txt: 352847 bytes, checksum: 4351a70b127ee05070c1b0e47ae7648c (MD5)\nGoncalves_grad.sunysb_0771E_11045.pdf: 9441512 bytes, checksum: b464b60b2a3519d4d79b5dccc85e94c1 (MD5)\n  Previous issue date:    1"},{"label":"dcterms.publisher","value":"The Graduate School, Stony Brook University: Stony Brook, NY."},{"label":"dcterms.subject","value":"Biochemistry"},{"label":"dcterms.title","value":"Correlating Structure and Function in Class A \nGPCRs"},{"label":"dcterms.type","value":"Dissertation"},{"label":"dc.type","value":"Dissertation"}],"description":"This manifest was generated dynamically","viewingDirection":"left-to-right","sequences":[{"@type":"sc:Sequence","canvases":[{"@id":"https://repo.library.stonybrook.edu/cantaloupe/iiif/2/canvas/page-1.json","@type":"sc:Canvas","label":"Page 1","height":1650,"width":1275,"images":[{"@type":"oa:Annotation","motivation":"sc:painting","resource":{"@id":"https://repo.library.stonybrook.edu/cantaloupe/iiif/2/43%2F40%2F48%2F43404887233591934736434216264876282399/full/full/0/default.jpg","@type":"dctypes:Image","format":"image/jpeg","height":1650,"width":1275,"service":{"@context":"http://iiif.io/api/image/2/context.json","@id":"https://repo.library.stonybrook.edu/cantaloupe/iiif/2/43%2F40%2F48%2F43404887233591934736434216264876282399","profile":"http://iiif.io/api/image/2/level2.json"}},"on":"https://repo.library.stonybrook.edu/cantaloupe/iiif/2/canvas/page-1.json"}]}]}]}