title: Quantitative and dynamic analysis of PTEN phosphorylation by NMR.
abstract: The dual lipid and protein phosphatase PTEN is a tumor suppressor controlling key biological processes, such as cell growth, proliferation and neuro-survival. Its activity and intracellular trafficking is finely regulated notably by multi-site phosphorylation of its C-terminal tail. The reversible and highly dynamic character of these regulatory events confers a temporal dimension to the cell for triggering crucial decisions. In this review, we describe how a recently developed time-resolved NMR spectroscopy approach unveils the dynamic establishment of the phosphorylation events of PTEN C-terminal tail controlled by CK2 and GSK3β kinases. Two cascades of reactions have been identified, in vitro and in extracts of human neuroblastoma cells. They are triggered independently on two nearby clusters of sites (S380-S385 and S361-S370) and occur on different timescales. In each cascade, the reactions follow an ordered model with a distributive kinetic mechanism. The vision of these cascades as two delay timers activating distinct or time-delayed regulatory responses gives a temporal dimension on PTEN regulation and is discussed in relation to the known functional roles of each cluster.
authors.affiliation: Institut Pasteur, Unité de Résonance Magnétique Nucléaire des Biomolécules, Département de Biologie Structurale et Chimie, F-75015 Paris, France; CNRS, UMR3528, F-75015 Paris, France. Electronic address: florence.cordier@pasteur.fr.
authors.forename: Florence
authors.initials: F
authors.lastname: Cordier
authors.affiliation: Institut Pasteur, Unité de Résonance Magnétique Nucléaire des Biomolécules, Département de Biologie Structurale et Chimie, F-75015 Paris, France; CNRS, UMR3528, F-75015 Paris, France.
authors.forename: Alain
authors.initials: A
authors.lastname: Chaffotte
authors.affiliation: Institut Pasteur, Unité de Résonance Magnétique Nucléaire des Biomolécules, Département de Biologie Structurale et Chimie, F-75015 Paris, France; CNRS, UMR3528, F-75015 Paris, France. Electronic address: nicolas.wolff@pasteur.fr.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Wolff
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D045744:Cell Line, Tumor; D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008969:Molecular Sequence Data; D051059:PTEN Phosphohydrolase / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation / Q000502:physiology; D025521:Tumor Suppressor Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*
chemical_list: D025521:Tumor Suppressor Proteins; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human
keywords: Distributive mechanism; Multi-site phosphorylation; Ordered reactions; PTEN; Phosphorylation kinetics; Time-resolved NMR spectroscopy