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Method Article
Purification and Liposome Reconstitution of a bacterial multi-transmembrane domain protein for detection of autokinase activity in response to signals
Vanessa Sperandio
Sperandio lab
Corresponding Author
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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
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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)
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This is a preprint. It has not completed peer review.
Method Article
Purification and Liposome Reconstitution of a bacterial multi-transmembrane domain protein for detection of autokinase activity in response to signals
Vanessa Sperandio
Sperandio lab
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
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).
Associated Publications
Fucose sensing regulates bacterial intestinal colonization
by Alline R. Pacheco, Meredith M. Curtis, Jennifer M. Ritchie, +4
Nature (27 November, 2012)