title: Detecting adenosine triphosphate in the pericellular space.
abstract: Release of adenosine triphosphate (ATP) into the extracellular space occurs in response to a multiplicity of physiological and pathological stimuli in virtually all cells and tissues. A role for extracellular ATP has been identified in processes as different as neurotransmission, endocrine and exocrine secretion, smooth muscle contraction, bone metabolism, cell proliferation, immunity and inflammation. However, ATP measurement in the extracellular space has proved a daunting task until recently. To tackle this challenge, some years ago, we designed and engineered a novel luciferase probe targeted to and expressed on the outer aspect of the plasma membrane. This novel probe was constructed by appending to firefly luciferase the N-terminal leader sequence and the C-terminal glycophosphatidylinositol anchor of the folate receptor. This chimeric protein, named plasma membrane luciferase, is targeted and localized to the outer side of the plasma membrane. With this probe, we have generated stably transfected HEK293 cell clones that act as an in vitro and in vivo sensor of the extracellular ATP concentration in several disease conditions, such as experimentally induced tumours and inflammation.
authors.affiliation: Department of Morphology, Surgery and Experimental Medicine, Section of General Pathology , University of Ferrara , Via Borsari 46, Ferrara 44121 , Italy.
authors.forename: Simonetta
authors.initials: S
authors.lastname: Falzoni
authors.forename: Giovanna
authors.initials: G
authors.lastname: Donvito
authors.forename: Francesco
authors.initials: F
authors.lastname: Di Virgilio
keywords: bioluminescence; cancer; extracellular adenosine triphosphate; inflammation; luciferase