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Title 

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

Authors 

Eun-Ju JeongKyoungsook ParkHyou-Arm JoungChang-Soo LeeD W SeolBong Hyun ChungMoonil Kim

Publisher 

Springer Verlag (Germany)

Issue Date 

2007

Citation 

Biotechnology Letters, vol. 29, no. 5, pp. 797-802

Keywords 

conformational changefluorescence complementation assayglucose-induced fithexokinase IIsplit EGFPbioassayfluorescenceglucose

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.

ISSN 

0141-5492

Link 

http://dx.doi.org/10.1007/s10529-007-9313-x

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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