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New page: left|200px<br /><applet load="2okw" size="350" color="white" frame="true" align="right" spinBox="true" caption="2okw, resolution 1.9Å" /> '''A non-invasive GFP-ba...
 
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[[Image:2okw.jpg|left|200px]]<br /><applet load="2okw" size="350" color="white" frame="true" align="right" spinBox="true"
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'''A non-invasive GFP-based biosensor for mercury ions'''<br />


==Overview==
==A non-invasive GFP-based biosensor for mercury ions==
Atomic resolution structures of proteins indicate that the core is, typically well packed, suggesting a densely connected network of, interactions between amino acid residues. The combinatorial complexity of, energetic interactions in such a network could be enormous, a problem that, limits our ability to relate structure and function. Here, we report a, case study of the complexity of amino acid interactions in a localized, region within the core of the GFP, a particularly stable and tightly, packed molecule. Mutations at three sites within the chromophore-binding, pocket display an overlapping pattern of conformational change and are, thermodynamically coupled, seemingly consistent with the dense network, model. However, crystallographic and energetic analyses of coupling, between mutations paint a different picture; pairs of mutations couple, through independent "hotspots" in the region of structural overlap. The, data indicate that, even in highly stable proteins, the core contains, sufficient plasticity in packing to uncouple high-order energetic, interactions of residues, a property that is likely general in proteins.
<StructureSection load='2okw' size='340' side='right'caption='[[2okw]], [[Resolution|resolution]] 1.90&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[2okw]] is a 6 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=2OKW OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2OKW FirstGlance]. <br>
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.9&#8491;</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CRO:{2-[(1R,2R)-1-AMINO-2-HYDROXYPROPYL]-4-(4-HYDROXYBENZYLIDENE)-5-OXO-4,5-DIHYDRO-1H-IMIDAZOL-1-YL}ACETIC+ACID'>CRO</scene></td></tr>
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=2okw FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2okw OCA], [https://pdbe.org/2okw PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2okw RCSB], [https://www.ebi.ac.uk/pdbsum/2okw PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2okw ProSAT]</span></td></tr>
</table>
== Function ==
[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.
== Evolutionary Conservation ==
[[Image:Consurf_key_small.gif|200px|right]]
Check<jmol>
  <jmolCheckbox>
    <scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/ok/2okw_consurf.spt"</scriptWhenChecked>
    <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview03.spt</scriptWhenUnchecked>
    <text>to colour the structure by Evolutionary Conservation</text>
  </jmolCheckbox>
</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=2okw ConSurf].
<div style="clear:both"></div>
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
Mercury is a ubiquitous pollutant that when absorbed is extremely toxic to a wide variety of biochemical processes. Mercury (II) is a strong, "invisible" poison that is rapidly absorbed by tissues of the intestinal tract, kidneys, and liver upon ingestion. In this study, a novel fluorescence-based biosensor is presented that allows for the direct monitoring of the uptake and distribution of the metal under noninvasive in vivo conditions. With the introduction of a cysteine residue at position 205, located in close proximity to the chromophore, the green fluorescent protein (GFP) from Aequorea victoria was converted into a highly specific biosensor for this metal ion. The mutant protein exhibits a dramatic absorbance and fluorescence change upon mercuration at neutral pH. Absorbance and fluorescence properties with respect to the metal concentration exhibit sigmoidal binding behavior with a detection limit in the low nanomolar range. Time-resolved binding studies indicate rapid subsecond binding of the metal to the protein. The crystal structures obtained of mutant eGFP205C indicate a possible access route of the metal into the core of the protein. To our knowledge, this engineered protein is a first example of a biosensor that allows for noninvasive and real-time imaging of mercury uptake in a living cell. A major advantage is that its expression can be genetically controlled in many organisms to enable unprecedented studies of tissue specific mercury uptake.


==About this Structure==
Design of a highly specific and noninvasive biosensor suitable for real-time in vivo imaging of mercury (II) uptake.,Chapleau RR, Blomberg R, Ford PC, Sagermann M Protein Sci. 2008 Apr;17(4):614-22. Epub 2008 Feb 27. PMID:18305194<ref>PMID:18305194</ref>
2OKW 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=2OKW OCA].


==Reference==
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
Local complexity of amino acid interactions in a protein core., Jain RK, Ranganathan R, Proc Natl Acad Sci U S A. 2004 Jan 6;101(1):111-6. Epub 2003 Dec 18. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=14684834 14684834]
</div>
<div class="pdbe-citations 2okw" style="background-color:#fffaf0;"></div>
 
==See Also==
*[[Green Fluorescent Protein 3D structures|Green Fluorescent Protein 3D structures]]
== References ==
<references/>
__TOC__
</StructureSection>
[[Category: Aequorea victoria]]
[[Category: Aequorea victoria]]
[[Category: Single protein]]
[[Category: Large Structures]]
[[Category: Blomberg, R.]]
[[Category: Blomberg R]]
[[Category: Chapleau, R.]]
[[Category: Chapleau RR]]
[[Category: Ford, P.]]
[[Category: Ford PC]]
[[Category: Sagermann, M.]]
[[Category: Sagermann M]]
[[Category: biosensor]]
[[Category: luminescent protein]]
[[Category: mercury]]
 
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