Methods in the Study of PTEN Structure: X-Ray Crystallography and Hydrogen Deuterium Exchange Mass Spectrometry. 2016

Glenn R Masson, and John E Burke, and Roger L Williams
Laboratory of Molecular Biology, Medical Research Council (MRC), Francis Crick Avenue, Cambridge Biomedical campus, Cambridge, CB2 0QH, UK.

Despite its small size and deceptively simple domain organization, PTEN remains a challenging structural target due to its N- and C-terminal intrinsically disordered segments, and the conformational heterogeneity caused by phosphorylation of its C terminus. Using hydrogen/deuterium exchange mass spectrometry (HDX-MS), it is possible to probe the conformational dynamics of the disordered termini, and also to determine how PTEN binds to lipid membranes. Here, we describe how to purify recombinant, homogenously dephosphorylated PTEN from a eukaryotic system for subsequent investigation with HDX-MS or crystallography.

UI MeSH Term Description Entries
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013058 Mass Spectrometry An analytical method used in determining the identity of a chemical based on its mass using mass analyzers/mass spectrometers. Mass Spectroscopy,Spectrometry, Mass,Spectroscopy, Mass,Spectrum Analysis, Mass,Analysis, Mass Spectrum,Mass Spectrum Analysis,Analyses, Mass Spectrum,Mass Spectrum Analyses,Spectrum Analyses, Mass
D041961 Deuterium Exchange Measurement A research technique to measure solvent exposed regions of molecules that is used to provide insight about PROTEIN CONFORMATION. Hydrogen-Deuterium Exchange Measurement,Deuterium Exchange Measurements,Exchange Measurement, Deuterium,Exchange Measurement, Hydrogen-Deuterium,Exchange Measurements, Deuterium,Exchange Measurements, Hydrogen-Deuterium,Hydrogen Deuterium Exchange Measurement,Hydrogen-Deuterium Exchange Measurements,Measurement, Deuterium Exchange,Measurement, Hydrogen-Deuterium Exchange,Measurements, Deuterium Exchange,Measurements, Hydrogen-Deuterium Exchange
D051059 PTEN Phosphohydrolase A lipid phosphatase that contains a C2 DOMAIN and acts on phosphatidylinositol-3,4,5-trisphosphate to regulate various SIGNAL TRANSDUCTION PATHWAYS. It modulates CELL GROWTH PROCESSES; CELL MIGRATION; and APOPTOSIS. Mutations in PTEN are associated with COWDEN DISEASE and PROTEUS SYNDROME as well as NEOPLASTIC CELL TRANSFORMATION. MMAC1 Protein,Mutated In Multiple Advanced Cancers 1 Protein,PTEN Phosphatase,PTEN Protein,PTEN Protein Phosphatase,Phosphatase and Tensin Homologue on Chromosome Ten Protein,Phosphatase, PTEN,Phosphatase, PTEN Protein,Phosphohydrolase, PTEN,Protein Phosphatase, PTEN
D018360 Crystallography, X-Ray The study of crystal structure using X-RAY DIFFRACTION techniques. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) X-Ray Crystallography,Crystallography, X Ray,Crystallography, Xray,X Ray Crystallography,Xray Crystallography,Crystallographies, X Ray,X Ray Crystallographies

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