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Purification and Liposome Reconstitution of a bacterial multi-transmembrane domain protein for detection of autokinase activity in response to signals

External protocol Created on 30 Apr 2014

Authors

Vanessa Sperandio

Summary

Autophosphorylation activity of histidine-kinases (HK) reconstituted into liposomes has been successfully employed in the Sperandio laboratory to study the response of HKs to chemical cues (1, 2). One of the challenges of performing autophosphorylation assays with membrane-bound HKs is the purification of functional proteins for liposome loading and reconstitution. In this protocol, we describe the liposome reconstitution of the multi-transmembrane domain HK FusK of Enterohemorrhagic Escherichia coli (EHEC) O157:H7, and in vitro autophosphorylation activity in response to L-fucose (5).

Further details

The protocol was published on Protocol Exchange on 27 November 2012. To see the entire protocol, click on the source link.

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