2f32: Difference between revisions

Revision as of 03:24, 4 May 2008

Xray crystal structure of lysozyme mutant L20/R63A liganded to ethylguanidinium

OverviewOverview

The binding of guanidinium ion has been shown to promote a large-scale translation of a tandemly duplicated helix in an engineered mutant of T4 lysozyme. The guanidinium ion acts as a surrogate for the guanidino group of an arginine side chain. Here we determine whether methyl- and ethylguanidinium provide better mimics. The results show that addition of the hydrophobic moieties to the ligand enhances the binding affinity concomitant with reduction in ligand solubility. Crystallographic analysis confirms that binding of the alternative ligands to the engineered site still drives the large-scale conformational change. Thermal analysis and NMR data show, in comparison to guanidinium, an increase in protein stability and in ligand affinity. This is presumably due to the successive increase in hydrophobicity in going from guanidinium to ethylguanidinium. A fluorescence-based optical method was developed to sense the ligand-triggered helix translation in solution. The results are a first step in the de novo design of a molecular switch that is not related to the normal function of the protein.

About this StructureAbout this Structure

2F32 is a Single protein structure of sequence from Enterobacteria phage t4. Full crystallographic information is available from OCA.

ReferenceReference

Guanidinium derivatives bind preferentially and trigger long-distance conformational changes in an engineered T4 lysozyme., Yousef MS, Bischoff N, Dyer CM, Baase WA, Matthews BW, Protein Sci. 2006 Apr;15(4):853-61. PMID:16600969 Page seeded by OCA on Sun May 4 03:24:23 2008

Proteopedia Page Contributors and Editors (what is this?)Proteopedia Page Contributors and Editors (what is this?)

OCA

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 [[Image:2f32.gif|left|200px]]
- {{Structure
+<!--
-|PDB= 2f32 |SIZE=350|CAPTION= <scene name='initialview01'>2f32</scene>, resolution 1.80&Aring;
+The line below this paragraph, containing "STRUCTURE_2f32", creates the "Structure Box" on the page.
-|SITE=
+You may change the PDB parameter (which sets the PDB file loaded into the applet)
-|LIGAND= <scene name='pdbligand=BME:BETA-MERCAPTOETHANOL'>BME</scene>, <scene name='pdbligand=EGD:N-ETHYLGUANIDINE'>EGD</scene>
+or the SCENE parameter (which sets the initial scene displayed when the page is loaded),
-|ACTIVITY= <span class='plainlinks'>[http://en.wikipedia.org/wiki/Lysozyme Lysozyme], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.17 3.2.1.17] </span>
+or leave the SCENE parameter empty for the default display.
-|GENE= E ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=10665 Enterobacteria phage T4])
+-->
-|DOMAIN=
+{{STRUCTURE_2f32|  PDB=2f32  |  SCENE=  }}
-|RELATEDENTRY=[[2f2q|2F2Q]]
-|RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2f32 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2f32 OCA], [http://www.ebi.ac.uk/pdbsum/2f32 PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=2f32 RCSB]</span>
-}}
 '''Xray crystal structure of lysozyme mutant L20/R63A liganded to ethylguanidinium'''
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 [[Category: Matthews, B W.]]
 [[Category: Yousef, M S.]]
-[[Category: molecular switch]]
+[[Category: Molecular switch]]
-[[Category: nano-bitechnology]]
+[[Category: Nano-bitechnology]]
-[[Category: protein design]]
+[[Category: Protein design]]
-[[Category: protein engineering]]
+[[Category: Protein engineering]]
-[[Category: t4 lysozyme]]
+[[Category: T4 lysozyme]]
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun May  4 03:24:23 2008''
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Mar 31 02:57:13 2008''