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2h3h

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{{STRUCTURE_2h3h| PDB=2h3h | SCENE= }}
{{STRUCTURE_2h3h| PDB=2h3h | SCENE= }}
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'''Crystal structure of the liganded form of Thermotoga maritima glucose binding protein'''
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===Crystal structure of the liganded form of Thermotoga maritima glucose binding protein===
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==Overview==
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We report the design and engineering of a robust, reagentless fluorescent glucose biosensor based on the periplasmic glucose-binding protein obtained from Thermotoga maritima (tmGBP). The gene for this protein was cloned from genomic DNA and overexpressed in Escherichia coli, the identity of its cognate sugar was confirmed, ligand binding was studied, and the structure of its glucose complex was solved to 1.7 Angstrom resolution by X-ray crystallography. TmGBP is specific for glucose and exhibits high thermostability (midpoint of thermal denaturation is 119 +/- 1 degrees C and 144 +/- 2 degrees C in the absence and presence of 1 mM glucose, respectively). A series of fluorescent conjugates was constructed by coupling single, environmentally sensitive fluorophores to unique cysteines introduced by site-specific mutagenesis at positions predicted to be responsive to ligand-induced conformational changes based on the structure. These conjugates were screened to identify engineered tmGBPs that function as reagentless fluorescent glucose biosensors. The Y13C*Cy5 conjugate is bright, gives a large response to glucose over concentration ranges appropriate for in vivo monitoring of blood glucose levels (1-30 mM), and can be immobilized in an orientation-specific manner in microtiter plates to give a reversible response to glucose. The immobilized protein retains its response after long-term storage at room temperature.
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{{ABSTRACT_PUBMED_17766373}}
==About this Structure==
==About this Structure==
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[[Category: Glucose binding protein]]
[[Category: Glucose binding protein]]
[[Category: Periplasmic binding protein]]
[[Category: Periplasmic binding protein]]
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Revision as of 22:31, 27 July 2008

Image:2h3h.png

Template:STRUCTURE 2h3h

Crystal structure of the liganded form of Thermotoga maritima glucose binding protein

Template:ABSTRACT PUBMED 17766373

About this Structure

2H3H is a Single protein structure of sequence from Thermotoga maritima. Full crystallographic information is available from OCA.

Reference

Structure-based design of robust glucose biosensors using a Thermotoga maritima periplasmic glucose-binding protein., Tian Y, Cuneo MJ, Changela A, Hocker B, Beese LS, Hellinga HW, Protein Sci. 2007 Oct;16(10):2240-50. Epub 2007 Aug 31. PMID:17766373

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