2hgd
From Proteopedia
| Line 1: | Line 1: | ||
| - | [[Image:2hgd.gif|left|200px]] | + | [[Image:2hgd.gif|left|200px]] |
| - | + | ||
| - | '''Structure of S65A Y66F GFP variant with an oxidized chromophore''' | + | {{Structure |
| + | |PDB= 2hgd |SIZE=350|CAPTION= <scene name='initialview01'>2hgd</scene>, resolution 1.60Å | ||
| + | |SITE= | ||
| + | |LIGAND= | ||
| + | |ACTIVITY= | ||
| + | |GENE= GFP ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=6100 Aequorea victoria]) | ||
| + | }} | ||
| + | |||
| + | '''Structure of S65A Y66F GFP variant with an oxidized chromophore''' | ||
| + | |||
==Overview== | ==Overview== | ||
| Line 7: | Line 16: | ||
==About this Structure== | ==About this Structure== | ||
| - | 2HGD is a [ | + | 2HGD is a [[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=2HGD OCA]. |
==Reference== | ==Reference== | ||
| - | The case of the missing ring: radical cleavage of a carbon-carbon bond and implications for GFP chromophore biosynthesis., Barondeau DP, Kassmann CJ, Tainer JA, Getzoff ED, J Am Chem Soc. 2007 Mar 21;129(11):3118-26. Epub 2007 Feb 28. PMID:[http:// | + | The case of the missing ring: radical cleavage of a carbon-carbon bond and implications for GFP chromophore biosynthesis., Barondeau DP, Kassmann CJ, Tainer JA, Getzoff ED, J Am Chem Soc. 2007 Mar 21;129(11):3118-26. Epub 2007 Feb 28. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/17326633 17326633] |
[[Category: Aequorea victoria]] | [[Category: Aequorea victoria]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| Line 23: | Line 32: | ||
[[Category: post-translational modification]] | [[Category: post-translational modification]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 17:16:42 2008'' |
Revision as of 15:16, 20 March 2008
| |||||||
| , resolution 1.60Å | |||||||
|---|---|---|---|---|---|---|---|
| Gene: | GFP (Aequorea victoria) | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
Structure of S65A Y66F GFP variant with an oxidized chromophore
Overview
The green fluorescent protein (GFP) creates its fluorophore by promoting spontaneous peptide backbone cyclization and amino acid oxidation chemistry on its own Ser65, Tyr66, Gly67 tripeptide sequence. Here we use high-resolution crystallography and mutational analyses to characterize GFP variants that undergo backbone cyclization followed by either anticipated chromophore synthesis via Y66F Calpha-Cbeta double-bond formation or unprecedented loss of a Y66F benzyl moiety via Calpha-Cbeta bond cleavage. We discovered a Y66F cleavage variant that subsequently incorporates an oxygen atom, likely from molecular oxygen, at the Y66 Calpha position. The post-translational products identified from these Y66F GFP structures support a common intermediate that partitions between Calpha-Cbeta oxidation and homolytic cleavage pathways. Our data indicate that Glu222 is the branchpoint control for this partitioning step and also influences subsequent oxygen incorporation reactions. From these results, we propose mechanisms for Y66F Calpha-Cbeta cleavage, oxygen incorporation, and chromophore biosynthesis with shared features that include radical chemistry. By revealing how GFP and RFP protein environments steer chemistry to favor fluorophore biosynthesis and disfavor alternative reactivity, we identify strategies for protein design. The proposed, common, one-electron oxidized, radical intermediate for post-translation modifications in the GFP family has general implications for how proteins drive and control spontaneous post-translational chemical modifications in the absence of metal ions.
About this Structure
2HGD is a Single protein structure of sequence from Aequorea victoria. Full crystallographic information is available from OCA.
Reference
The case of the missing ring: radical cleavage of a carbon-carbon bond and implications for GFP chromophore biosynthesis., Barondeau DP, Kassmann CJ, Tainer JA, Getzoff ED, J Am Chem Soc. 2007 Mar 21;129(11):3118-26. Epub 2007 Feb 28. PMID:17326633
Page seeded by OCA on Thu Mar 20 17:16:42 2008
