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Method Article
Vanessa Sperandio
Sperandio lab
Corresponding Author
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This work is licensed under a CC BY-NC 3.0 License. Read Full License
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).
Keywords
trans-membrane proteins, liposomes histidine kinases
no financial conflicts of interest
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Associated Publications
Fucose sensing regulates bacterial intestinal colonization
by Alline R. Pacheco, Meredith M. Curtis, Jennifer M. Ritchie, +4
Nature (27 November, 2012)
Method Article
Vanessa Sperandio
Sperandio lab
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 27 Nov, 2012
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
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).
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Associated Publications
Fucose sensing regulates bacterial intestinal colonization
by Alline R. Pacheco, Meredith M. Curtis, Jennifer M. Ritchie, +4
Nature (27 November, 2012)