2g6e
From Proteopedia
(New page: 200px<br /><applet load="2g6e" size="450" color="white" frame="true" align="right" spinBox="true" caption="2g6e, resolution 1.30Å" /> '''Structure of cyclize...) |
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| - | [[Image:2g6e.gif|left|200px]]<br /><applet load="2g6e" size=" | + | [[Image:2g6e.gif|left|200px]]<br /><applet load="2g6e" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="2g6e, resolution 1.30Å" /> | caption="2g6e, resolution 1.30Å" /> | ||
'''Structure of cyclized F64L S65A Y66S GFP variant'''<br /> | '''Structure of cyclized F64L S65A Y66S GFP variant'''<br /> | ||
==Overview== | ==Overview== | ||
| - | The green fluorescent protein (GFP) creates a fluorophore out of three | + | The green fluorescent protein (GFP) creates a fluorophore out of three sequential amino acids by promoting spontaneous posttranslational modifications. Here, we use high-resolution crystallography to characterize GFP variants that not only undergo peptide backbone cyclization but additional denaturation-induced peptide backbone fragmentation, native peptide hydrolysis, and decarboxylation reactions. Our analyses indicate that architectural features that favor GFP peptide cyclization also drive peptide hydrolysis. These results are relevant for the maturation pathways of GFP homologues, such as the kindling fluorescent protein and the Kaede protein, which use backbone cleavage to red-shift the spectral properties of their chromophores. We further propose a photochemical mechanism for the decarboxylation reaction, supporting a role for the GFP protein environment in facilitating radical formation and one-electron chemistry, which may be important in activating oxygen for the oxidation step of chromophore biosynthesis. Together, our results characterize GFP posttranslational modification chemistry with implications for the energetic landscape of backbone cyclization and subsequent reactions, and for the rational design of predetermined spontaneous backbone cyclization and cleavage reactions. |
==About this Structure== | ==About this Structure== | ||
| - | 2G6E is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Aequorea_victoria Aequorea victoria] with MG as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http:// | + | 2G6E is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Aequorea_victoria Aequorea victoria] with <scene name='pdbligand=MG:'>MG</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2G6E OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Aequorea victoria]] | [[Category: Aequorea victoria]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Barondeau, D | + | [[Category: Barondeau, D P.]] |
[[Category: MG]] | [[Category: MG]] | ||
[[Category: biosynthesis]] | [[Category: biosynthesis]] | ||
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[[Category: post-translational modification]] | [[Category: post-translational modification]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:28:38 2008'' |
Revision as of 15:28, 21 February 2008
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Structure of cyclized F64L S65A Y66S GFP variant
Overview
The green fluorescent protein (GFP) creates a fluorophore out of three sequential amino acids by promoting spontaneous posttranslational modifications. Here, we use high-resolution crystallography to characterize GFP variants that not only undergo peptide backbone cyclization but additional denaturation-induced peptide backbone fragmentation, native peptide hydrolysis, and decarboxylation reactions. Our analyses indicate that architectural features that favor GFP peptide cyclization also drive peptide hydrolysis. These results are relevant for the maturation pathways of GFP homologues, such as the kindling fluorescent protein and the Kaede protein, which use backbone cleavage to red-shift the spectral properties of their chromophores. We further propose a photochemical mechanism for the decarboxylation reaction, supporting a role for the GFP protein environment in facilitating radical formation and one-electron chemistry, which may be important in activating oxygen for the oxidation step of chromophore biosynthesis. Together, our results characterize GFP posttranslational modification chemistry with implications for the energetic landscape of backbone cyclization and subsequent reactions, and for the rational design of predetermined spontaneous backbone cyclization and cleavage reactions.
About this Structure
2G6E is a Single protein structure of sequence from Aequorea victoria with as ligand. Full crystallographic information is available from OCA.
Reference
Understanding GFP posttranslational chemistry: structures of designed variants that achieve backbone fragmentation, hydrolysis, and decarboxylation., Barondeau DP, Kassmann CJ, Tainer JA, Getzoff ED, J Am Chem Soc. 2006 Apr 12;128(14):4685-93. PMID:16594705
Page seeded by OCA on Thu Feb 21 17:28:38 2008
