1myw
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(New page: 200px<br /><applet load="1myw" size="450" color="white" frame="true" align="right" spinBox="true" caption="1myw, resolution 2.20Å" /> '''CRYSTAL STRUCTURE OF...)
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Revision as of 19:42, 20 November 2007
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CRYSTAL STRUCTURE OF A YELLOW FLUORESCENT PROTEIN WITH IMPROVED MATURATION AND REDUCED ENVIRONMENTAL SENSITIVITY
Overview
Yellow emission variants of green fluorescent protein (GFP) have been, found useful in a variety of applications in biological systems due to, their red-shifted emission spectrum and sensitivity to environmental, parameters, such as pH and ionic strength. However, slow maturation, properties and new requirements for more intense fluorescence necessitated, further mutagenesis studies of these proteins. Venus, a new variant with, improved maturation and brightness, as well as reduced environmental, dependence, was recently developed by introducing five mutations into the, well characterized variant, enhanced yellow fluorescent protein (EYFP). In, this paper, we present the crystal structure of Venus at 2.2 A resolution, which enabled us to correlate its novel features with these mutation, points. The rearrangement of several side chains near the chromophore, initiated by the F46L mutation, was found to improve maturation at 37, degrees C by removing steric and energetic constraints, which may hinder, folding of the polypeptide chain, and by accelerating the oxidation of the, Calpha-Cbeta bond of Tyr(66) during chromophore formation. M153T, V163A, and S175G were also found to improve the rate of maturation by creating, regions of greater flexibility. F64L induced large conformational changes, in the molecule, leading to the removal of halide sensitivity by, preventing ion access to the binding site.
About this Structure
1MYW is a Single protein structure of sequence from Aequorea victoria. Full crystallographic information is available from OCA.
Reference
Crystal structure of venus, a yellow fluorescent protein with improved maturation and reduced environmental sensitivity., Rekas A, Alattia JR, Nagai T, Miyawaki A, Ikura M, J Biol Chem. 2002 Dec 27;277(52):50573-8. Epub 2002 Oct 4. PMID:12370172
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