Detection of glucose-induced conformational change in hexokinase II using fluorescence complementation assay

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Title
Detection of glucose-induced conformational change in hexokinase II using fluorescence complementation assay
Author(s)
Eun-Ju Jeong; Kyoungsook Park; Hyou-Arm Joung; Chang-Soo Lee; D W Seol; Bong Hyun Chung; Moonil Kim
Bibliographic Citation
Biotechnology Letters, vol. 29, no. 5, pp. 797-802
Publication Year
2007
Abstract
Conformational changes in hexokinase are induced by its binding to glucose, thus providing an excellent example of an 'induced fit' model. To observe glucose-induced fluorescence restoration in hexokinase II using split-enhanced, green fluorescent protein (EGFP) in a process involving the reconstitution of split EGFP, E. coli cells expressing the chimeric NEGFP:HXK:CEGFP recombinant protein were treated with glucose and visualized via fluorescence read-outs. The reconstituted EGFP generated a strong fluorescence upon glucose stimulation of the bacteria. Moreover, the fluorescence intensity became stronger with increasing glucose up to 10 mM, with a maximum being observed after 60 min in a time- and concentration-dependent manner. Conformational changes associated with glucose-induced fit in human hexokinase II can thus be monitored successfully in vivo via fluorescence reconstitution assays, coupled with a quick and easy fluorescent read-out protocol.
Keyword
Conformational changeFluorescence complementation assayGlucose-induced fitHexokinase IISplit EGFP
ISSN
0141-5492
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s10529-007-9313-x
Type
Article
Appears in Collections:
Division of Research on National Challenges > Bionanotechnology Research Center > 1. Journal Articles
Critical Diseases Diagnostics Convergence Research Center > 1. Journal Articles
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