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1yhg

From Proteopedia

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[[Image:1yhg.gif|left|200px]]<br /><applet load="1yhg" size="350" color="white" frame="true" align="right" spinBox="true"
 
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caption="1yhg, resolution 2.500&Aring;" />
 
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'''Uncyclized precursor structure of S65G Y66S V68G GFP variant'''<br />
 
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==Overview==
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==Uncyclized precursor structure of S65G Y66S V68G GFP variant==
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The Aequorea victoria green fluorescent protein (GFP) undergoes a remarkable post-translational modification to create a chromophore out of its component amino acids S65, Y66, and G67. Here, we describe mutational experiments in GFP designed to convert this chromophore into a 4-methylidene-imidazole-5-one (MIO) moiety similar to the post-translational active-site electrophile of histidine ammonia lyase (HAL). Crystallographic structures of GFP variant S65A Y66S (GFPhal) and of four additional related site-directed mutants reveal an aromatic MIO moiety and mechanistic details of GFP chromophore formation and MIO biosynthesis. Specifically, the GFP scaffold promotes backbone cyclization by (1) favoring nucleophilic attack by close proximity alignment of the G67 amide lone pair with the pi orbital of the residue 65 carbonyl and (2) removing enthalpic barriers by eliminating inhibitory main-chain hydrogen bonds in the precursor state. GFP R96 appears to induce structural rearrangements important in aligning the molecular orbitals for ring cyclization, favor G67 nitrogen deprotonation through electrostatic interactions with the Y66 carbonyl, and stabilize the reduced enolate intermediate. Our structures and analysis also highlight negative design features of the wild-type GFP architecture, which favor chromophore formation by destabilizing alternative conformations of the chromophore tripeptide. By providing a molecular basis for understanding and controlling the driving force and protein chemistry of chromophore creation, this research has implications for expansion of the genetic code through engineering of modified amino acids.
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<StructureSection load='1yhg' size='340' side='right'caption='[[1yhg]], [[Resolution|resolution]] 2.50&Aring;' scene=''>
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== Structural highlights ==
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<table><tr><td colspan='2'>[[1yhg]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Aequorea_victoria Aequorea victoria]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1YHG OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1YHG FirstGlance]. <br>
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</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.5&#8491;</td></tr>
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<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=1yhg FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1yhg OCA], [https://pdbe.org/1yhg PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1yhg RCSB], [https://www.ebi.ac.uk/pdbsum/1yhg PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1yhg ProSAT]</span></td></tr>
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</table>
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== Function ==
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[https://www.uniprot.org/uniprot/GFP_AEQVI GFP_AEQVI] Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin.
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== Evolutionary Conservation ==
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[[Image:Consurf_key_small.gif|200px|right]]
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Check<jmol>
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<jmolCheckbox>
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<scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/yh/1yhg_consurf.spt"</scriptWhenChecked>
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<scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked>
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<text>to colour the structure by Evolutionary Conservation</text>
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</jmolCheckbox>
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</jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=1yhg ConSurf].
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<div style="clear:both"></div>
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==About this Structure==
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==See Also==
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1YHG is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Aequorea_victoria Aequorea victoria]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1YHG OCA].
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*[[Green Fluorescent Protein 3D structures|Green Fluorescent Protein 3D structures]]
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__TOC__
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==Reference==
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</StructureSection>
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Understanding GFP chromophore biosynthesis: controlling backbone cyclization and modifying post-translational chemistry., Barondeau DP, Kassmann CJ, Tainer JA, Getzoff ED, Biochemistry. 2005 Feb 15;44(6):1960-70. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=15697221 15697221]
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[[Category: Aequorea victoria]]
[[Category: Aequorea victoria]]
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[[Category: Single protein]]
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[[Category: Large Structures]]
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[[Category: Barondeau, D P.]]
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[[Category: Barondeau DP]]
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[[Category: Getzoff, E D.]]
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[[Category: Getzoff ED]]
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[[Category: Kassmann, C J.]]
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[[Category: Kassmann CJ]]
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[[Category: Tainer, J A.]]
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[[Category: Tainer JA]]
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[[Category: chromophore uncyclized dimer]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:05:19 2008''
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Current revision

Uncyclized precursor structure of S65G Y66S V68G GFP variant

PDB ID 1yhg

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