Structural properties of AMP-activated protein kinase

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Author(s) Riek, Uwe, Scholz, Roland, Konarev, Peter, Rufer, Arne, Suter, Marianne, Nazabal, Alexis, Ringler, Philippe, Chami, Mohamed, Müller, Shirley A., Neumann, Dietbert, Forstner, Michael, Hennig, Michael, Zenobi, Renato, Engel, Andreas, Svergun, Dmitri, Schlattner, Uwe, Wallimann, Theo
Publication Type Journal Items, Publication Status: Published
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Title Structural properties of AMP-activated protein kinase
Subtitle Dimerization, molecular shape, and changes upon ligand-binding
Author(s) Riek, Uwe
Scholz, Roland
Konarev, Peter
Rufer, Arne
Suter, Marianne
Nazabal, Alexis
Ringler, Philippe
Chami, Mohamed
Müller, Shirley A.
Neumann, Dietbert
Forstner, Michael
Hennig, Michael
Zenobi, Renato
Engel, Andreas
Svergun, Dmitri
Schlattner, Uwe
Wallimann, Theo
Journal or Series Title The journal of biological chemistry
Volume Number 283
Issue Number 26
Start Page 18331
End Page 18343
ISSN 0021-9258
1083-351X
Publisher The American Society for Biochemistry and Molecular Biology
Publication Place Bethesda
Publication Date 2008
Abstract Heterotrimeric AMP-activated protein kinase (AMPK) is crucial for energy homeostasis of eukaryotic cells and organisms. Here we report on (i) bacterial expression of untagged mammalian AMPK isoform combinations, all containing {gamma}1, (ii) an automated four-dimensional purification protocol, and (iii) biophysical characterization of AMPK heterotrimers by small angle x-ray scattering in solution (SAXS), transmission and scanning transmission electron microscopy (TEM, STEM), and mass spectrometry (MS). AMPK in solution at low concentrations (~1 mg/ml) largely consisted of individual heterotrimers in TEM analysis, revealed a precise 1:1:1 stoichiometry of the three subunits in MS, and behaved as an ideal solution in SAXS. At higher AMPK concentrations, SAXS revealed concentration-dependent, reversible dimerization of AMPK heterotrimers and formation of higher oligomers, also confirmed by STEM mass measurements. Single particle reconstruction and averaging by SAXS and TEM, respectively, revealed similar elongated, flat AMPK particles with protrusions and an indentation. In the lower AMPK concentration range, addition of AMP resulted in a significant decrease of the radius of gyration by ~5% in SAXS, which indicates a conformational switch in AMPK induced by ligand binding. We propose a structural model involving a ligand-induced relative movement of the kinase domain resulting in a more compact heterotrimer and a conformational change in the kinase domain that protects AMPK from dephosphorylation of Thr172, thus positively affecting AMPK activity.
DOI 10.1074/jbc.M708379200
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03430
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NEBIS System Number 000040237
Source Database ID PP-41353
WOS-000256949200061
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@article{Rk2008,
  author = "Riek, Uwe and Scholz, Roland and Konarev, Peter and Rufer, Arne and Suter, Marianne and Nazabal, Alexis and Ringler, Philippe and Chami, Mohamed and M{\"{u}}ller, Shirley A. and Neumann, Dietbert and Forstner, Michael and Hennig, Michael and Zenobi, Renato and Engel, Andreas and Svergun, Dmitri and Schlattner, Uwe and Wallimann, Theo",
  title = "{S}tructural properties of {A}{M}{P}-activated protein kinase: {D}imerization, molecular shape, and changes upon ligand-binding",
  journal = "The journal of biological chemistry",
  year = 2008,
  volume = "283",
  number = "26",
  pages = "18331--18343",
}


E-Citations record created: Thu, 01 Apr 2010, 22:57:03 CET