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| - | [[Image:2p0x.jpg|left|200px]] | |
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| - | <!--
| + | ==solution structure of a non-biological ATP-binding protein== |
| - | The line below this paragraph, containing "STRUCTURE_2p0x", creates the "Structure Box" on the page.
| + | <StructureSection load='2p0x' size='340' side='right'caption='[[2p0x]]' scene=''> |
| - | You may change the PDB parameter (which sets the PDB file loaded into the applet)
| + | == Structural highlights == |
| - | or the SCENE parameter (which sets the initial scene displayed when the page is loaded),
| + | <table><tr><td colspan='2'>[[2p0x]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Synthetic_gene Synthetic gene]. Full experimental information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2P0X OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2P0X FirstGlance]. <br> |
| - | or leave the SCENE parameter empty for the default display.
| + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">Solution NMR</td></tr> |
| - | -->
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ATP:ADENOSINE-5-TRIPHOSPHATE'>ATP</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr> |
| - | {{STRUCTURE_2p0x| PDB=2p0x | SCENE= }}
| + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=2p0x FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2p0x OCA], [https://pdbe.org/2p0x PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2p0x RCSB], [https://www.ebi.ac.uk/pdbsum/2p0x PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2p0x ProSAT]</span></td></tr> |
| - | | + | </table> |
| - | '''solution structure of a non-biological ATP-binding protein'''
| + | __TOC__ |
| - | | + | </StructureSection> |
| - | | + | [[Category: Large Structures]] |
| - | ==Overview== | + | [[Category: Synthetic gene]] |
| - | We present a structural and functional analysis of the evolutionary optimization of a non-biological protein derived from a library of random amino acid sequences. A series of previously described in vitro selection experiments transformed a low-affinity ancestral sequence into a stably folded, high affinity ATP binding protein structure. While the evolutionarily optimized protein differs from its ancestral sequence through the accumulation of 12 amino acid mutations, the means by which those mutations enhance the stability and functionality of the protein were not well understood. We used a combination of mutagenesis, biochemistry, and NMR spectroscopy to investigate the structural and functional significance of each mutation. We solved the three-dimensional structure of the folding optimized protein by solution NMR, which revealed a fourth strand of the beta-sheet of the alpha/beta-fold that was not observed in an earlier crystallographic analysis of a less stable version of the protein. The structural rigidity of the newly identified beta-strand was confirmed by T1, T2, and heteronuclear nuclear Overhauser enhancement (NOE) measurements. Biochemical experiments were used to examine point mutations that revert the optimized protein back to the ancestral residue at each of the 12 sites. A combination of structural and functional data was then used to interpret the significance of each amino acid mutation. The enhanced ATP affinity was largely due to the emergence of a patch of positive charge density on the protein surface, while the increased solubility resulted from several mutations that increased the hydrophilicity of the protein surface, thereby decreasing protein aggregation. One mutation may stabilize the hydrophobic face of the beta-sheet.
| + | [[Category: Chaput JC]] |
| - | | + | [[Category: Mansy SS]] |
| - | ==About this Structure==
| + | [[Category: Szostak JW]] |
| - | Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2P0X OCA]. | + | |
| - | | + | |
| - | ==Reference== | + | |
| - | Structure and evolutionary analysis of a non-biological ATP-binding protein., Mansy SS, Zhang J, Kummerle R, Nilsson M, Chou JJ, Szostak JW, Chaput JC, J Mol Biol. 2007 Aug 10;371(2):501-13. Epub 2007 May 26. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/17583732 17583732]
| + | |
| - | [[Category: Chaput, J C.]] | + | |
| - | [[Category: Mansy, S S.]] | + | |
| - | [[Category: Szostak, J W.]] | + | |
| - | [[Category: Alpha/beta fold]] | + | |
| - | [[Category: De novo protein]] | + | |
| - | [[Category: Treble clef zinc binding motif]] | + | |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun May 4 12:06:11 2008''
| + | |
